Scientia Horticulturae 299 (2022) 111041

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

Effect of Azospirillum brasilense on the in vitro germination of Eustoma

grandiflorum (Raf.) Schinn.(Gentianaceae)
María Paula Santos a, b, Sergio Javier Martínez a, Mauro Enrique Yarte a, b,
Susana Margarita Carletti a, Ezequiel Enrique Larraburu a, b, *
a
Laboratorio de Cultivo de Tejidos Vegetales (CULTEV) – Departamento de Ciencias Básicas- Universidad Nacional de Luján, Luján (Bs As), Argentina
b
Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Argentina

A R T I C L E I N F O A B S T R A C T

Key words: Lisianthus (Eustoma grandiflorum (Raf) Shinn.) is an excellent ornamental cut variety because it has flowers with
Azospirillum brasilense Az39 good vase life. Seed propagation is difficult due to the long production time (it takes up to 100 days for trans­
Lisianthus planting) and the small size of its seeds. In vitro germination may produce healthy and homogeneous seedlings
In vitro germination
and may be used as a model system to study the effect of plant growth promoting rhizobacteria. Azospirillum
Biometrics parameters
brasilense is widely studied for its ability to colonize and promote various plant species growth, especially by
Mesophyll thickness, root aerenchyma
increasing root development and plant biomass. This work aimed to evaluate physiological, morphological and
anatomical aspects involved in vitro germination of E. grandiflorum inoculated with Azospirillum brasilense Az39.
Seeds of E. grandiflorum (F1 double big. Arena III Pink) were disinfected with 10% NaClO and culture in MS
medium. Inoculation of A. brasilense Az39 was performed with 106 and 107 cfu on seed surface at sowing. Non-
inoculated seeds were used as controls. Biofertilization generated earliness and higher germination percentages.
Furthermore, bacterization with 106 cfu significantly increased number of leaves, leaf area, fresh and dry weight
of seedlings, length and number of roots, diameter of the vascular bundle, leaves thickness and root thickness,
among other parameters. These results suggested that inoculation of lisianthus with A. brasilense Az39 could
improve seedlings and lead to a shorter time production.

Introduction
The production and marketing of flowers is an economic activity that
is practiced in almost 150 countries around the world. Cut flowers
constitute the main group within global floriculture trade and its pro­
duction has been growing around the world. The main producing and
consuming countries of cut flowers are the Netherlands in Europe, the
United States in North America and Japan in Asia. These countries are
shipping more than 50% of world trade (Yumbla-Orbes et al., 2017;
Wani et al., 2018).
Lisianthus Eustoma grandiflorum (Raf.) Schinn. (Gentianaceae) is
native from the humid grasslands of the southern United States and
northern Mexico (Fernández-Pavía and Trejo-Téllez, 2018). It is an
ornamental plant that stands out for its diversity of flower colours and
their good duration in the vase thus becoming an excellent cut variety
(Harbaugh, 2007). E. grandiflorum is propagated from seeds, which are
very small and difficult to grow. For this reason, seeds are commer­
cialized in pelleted form, coated with solid and inert materials that
modify their shape and size to facilitate handling (Bhatia and Sindhu,
2019). Required time for this species could be from 100 to 120 days from
transplant to harvest and is strongly influenced by the environment
(radiation and temperature) and the variety (Harbaugh, 2007). In
addition, the crop is attacked by thrips, caterpillars, leafminers,
Rhizoctonia sp., Fusarium sp. and Botrytis cinerea, among others that
affect the productive quality (Doğan et al., 2018). Finally, the produc­
tion of seedlings using conventional systems is usually not efficient,
obtaining heterogeneous individuals with reduced aerial and root
development (Barbaro et al., 2009). For these reasons, it is necessary to
find biotechnological tools to improve the propagation process.
In vitro techniques involves an aseptic environment, sterile nutrient
media and optimal conditions of light and temperature, which allow to
solve some inhibition problems during germination, increasing germi­
nation rates, reducing seedling loss due to contamination with phyto­
pathogens, and obtaining homogeneous seedlings (Hazarika, 2006;
George et al., 2008). Furthermore, it is possible to obtain a clonal pool of
seedlings under aseptic and controlled conditions, which can continue

* Corresponding author.

https://doi.org/10.1016/j.scienta.2022.111041
Received 27 August 2021; Received in revised form 7 January 2022; Accepted 5 March 2022
Available online 14 March 2022
0304-4238/© 2022 Elsevier B.V. All rights reserved.
M.P. Santos et al.
into micropropagation process or conventional cultivation.
Plant growth-promoting rhizobacteria (PGPR) produce beneficial
effects for the plant by direct or indirect mechanisms (Gupta et al.,
2015). Some mechanism include the production of phytohormones
(mainly indole-3-acetic acid); increasing nutrients availability, oxida­
tive enzyme activity and biological nitrogen fixation; decrease in the
incidence of diseases (due to the production of hydrocyanic acid, side­
rophores, antibiotics or competition for nutrients); improvement of
drought tolerance, salinity and toxicity to metals (Beneduzi et al., 2012;
Larraburu et al., 2016b). Within the PGPR group, the genus Azospirillum
is one of the most studied and used for inoculants formulation in the
world for its ability to fix atmospheric nitrogen and stimulate plant
growth (Hungria et al., 2010). The response of various crops to inocu­
lation with Azospirillum has been extensively studied, both in experi­
ments under controlled conditions and in the field (Pereg et al., 2016).
In most of the studies carried out under controlled conditions,
growth promotion by Azospirillum was clearly demonstrated (Spaepen
et al., 2009; Llorente and Larraburu, 2012; Gonzalez et al., 2015; Lar­
raburu et al., 2016a, b). Biofertilizers or inoculants appear as a useful
tool for the provision of nutrients to crops, with promising responses on
plant development, yield and health (Pereg et al., 2016).
Although PGPR in plant tissue culture has been demonstrated
increasing multiplication and rooting and explant quality in several
species (Orlikowska et al., 2017; Quambusch and Winkelmann, 2018),
the interaction between Lisianthus and PGPR is little explored. So the
aim of this work was to evaluate physiological, morphological and
anatomical aspects in in vitro germination of E. grandiflorum seeds
inoculated with A. brasilense Az39.
Material and methods
Bacterial strain and culture conditions
Azospirillum brasilense Az39 (strain deposited at the BPCV-IMyZA-
INTA Culture Collection, Castelar, Buenos Aires, Argentina) was used
for experiments.
Bacterial inoculum was grown in 250 mL Erlenmeyer flask with 150
mL liquid medium (Okon et al., 1977) at 32 ± 1 ◦ C for 72 h in an orbital
shaker Sontec™ at 140 rpm. The number of colony-forming units (cfu)
in each culture was determined in duplicate by dilution plate counts on
Congo Red (RC) medium (Rodríguez-Cáceres, 1982). Plates were
cultured at 37◦ C for 48 h.
In vitro germination of eustoma grandiflorum and bacterial inoculation
Pelleted seeds of E. grandiflorum (F1 double big. Arena III pink, TAKII
SEED) provided by the Floriculture Institute (INTA Castelar, Buenos
Aires, Argentina) were disinfected by immersion in sodium hypochlorite
(2.5 v/v of active chlorine) for 10 min. with stirring. Subsequently, the
seeds were rinsed 3 times with sterile distilled water. Each axenic seed
was grown in a flat-bottom glass tube (55 mL) with 15 mL of basic
medium Murashige and Skoog (1962) salts (MS) supplemented with
0.7% agar and pH 5.8. Inoculation was made on seed surface at sowing
tadding 0.1 mL A. brasilense Az39 complete inoculum (107 c.f.u per seed)
or its dilution 1:10 in sterile NaCl 0,85% (106 c.f.u per seed).
Non-inoculated treatments were used as control.
Cultures were incubated in a growth chamber at 25±2 ◦ C with
55–60% relative humidity under Phillips fluorescent daylight tubes (40
±4 μmol m − 2 s − 1) with a 16-h photoperiod.
Biometric parameters
The germination percentage was recorded at 10, 20 and 30 days after
inoculation. Fresh and dry weights of shoots and roots (samples were
dried at 60 ◦ C until constant weight), number of leaves and roots, main
shoot and root length were measured at 60 days. In addition, areas of
2
Scientia Horticulturae 299 (2022) 111041
scanned leaves were determined by ImageJ software (Drienovsky et al.,
2017).
Anatomical techniques
Leaves and roots from seedlings obtained at the end of the experi­
ments were collected and fixed in FAA (2:1:10:7 formalin: acetic acid:
ethanol 96 ◦ : distilled water). Samples were included in paraffin and cut
transversely (18–20 μm) using rotatory microtome MICRON HM325.
Then, sections were stained with safranin-fast green and mounted in
synthetic Canadian balsam (Biopack, Argentina), according to conven­
tional methodology (Zarlavsky, 2014). Observations and photographs
were realized using digital camera Megapixels attached to a light mi­
croscope XSZ 107B Leaf thickness, epidermis thickness, root cortex
thickness, rhizodermis thickness, root and central cylinder diameter, as
well as parenchymal cell size were determined. Picture capturing and
processing were performed using the TSview® software (Tucson Imag­
ing Technology Co.).
Experimental design and statistical analysis
Experiments were performed on the basis of a completely random­
ized design with three replicates per experiment, with at least 20 repe­
titions per treatment. Germination percentage was analyzed by
logarithmic regression analysis and Z-test for rate comparisons. Bio­
metric and anatomical parameters data were performed using a one way
ANOVA (analysis of variance) and multiple comparisons of means
(Tukey’s test). All statistical analyses were carried out using SPSS v 21.0
(SPSS Inc.).
Results
Seeds inoculated with 106 and 107 cfu of A. brasilense Az39 showed
early and higher germination rate (Fig. 1). Germination percentage was
low in inoculated seeds (5–10%) and null in control at ten days. Then,
bacterization increased the germination percentage around 93% at 20
days, and 68% at 30 days. No significant differences between both
inoculation treatments and similar regression curves were observed
(Fig. 1).
Inoculation of seed with A. brasilense Az39 106 cfu significantly
increased all biometric parameters (p ≤ 0.05) with respect to control and
seed inoculation with higher cfu (Fig. 2, Table 1). This treatment
increased leaf area by 300%, number of leaves by 59%, fresh shoot
weight by 2783%, dry shoot weight by 431%, fresh root weight by
5775%, dry shoot weight by 2455% and root length by 128% with
respect to control. In addition, A. brasilense Az39 107cfu significantly
increased only fresh shoot and root weight by 487% and 534%,
respectively (Fig. 2, Table 1).
Changes in foliar anatomy of lisianthus were observed in both
bacterization treatments with A. brasilense Az39, compared to non-
inoculated treatment (Fig. 3). Increments of 192 and 105% were
determined in total leaf thickness by inoculating with 106 and 107cfu
respectively, compared with the control treatment (Table 2). In this
sense, mesophyll thickness was affected in both inoculated treatments,
but particularly with 106 cfu of A. brasilense, which showed significantly
higher values (Table 2). Increases in the size of chlorenchyma cells also
were shown in the bacterization treatments, compared to control.
Furthermore, A. brasilense Az39 106 cfu treatment produced the greatest
effect increasing 149%, whereas 107 cfu of A. brasilense treatment only
increasing 27% (Table 2).
In addition, the thickness of the abaxial and adaxial epidermis were
significantly increased in both inoculated treatments with respect to the
control, although the highest values were exhibited by the 106 cfu
treatment (Table 2).
Regarding root anatomy, significant changes occurred in the inocu­
lated treatments (Fig. 4). Significant differences were observed in most
M.P. Santos et al. Scientia Horticulturae 299 (2022) 111041

Fig. 1. Effect of Azospirillum brasilense Az39 bacterization on in vitro germination percentage of Eustoma grandiflorum seeds during 30 days of culture. Control: non-
inoculated seedlings; A. brasilense Az39 106 cfu: inoculation at sowing with 1.3 106 cfu; A. brasilense Az39 107 cfu: inoculation at sowing with 1.3 107 cfu . Logaritmic
regresion curves, coefficient of determination and ecuations are in the same color of the each point serie .

Fig. 2. E. grandiflorum seedlings grown in vitro after 60 days of culture. (A), non-inoculated seedlingss (control); (B), inoculation at seeding with A. brasilense Az39
1.3 × 106 cfu of Az39; (C) inoculation at seeding with A. brasilense Az39 1.3 × 107 cfu.

of the evaluated parameters, being A. brasilense 106 cfu the treatment
that showed the largest significant increases (Table 3). No significant
differences were observed between control and seed inoculated with 107
cfu of A. brasilense on diameter of the central cylinder (Table 3). In
addition, roots obtained from inoculated seedlings with 107 cfu of
A. brasilense, exhibited significant aerenchyma with large intercellular
spaces in the bark. This feature is also observed in 106 cfu treatment but
with smaller intercellular spaces and larger parenchymal cells (Fig 4).
Discussion
Lisianthus seeds inoculation with A. brasilense Az39 generated sig­
nificant increases in seed germination percentage (52–57%), precocity,
biometrics parámeter and anatomical features, which is valuable to
obtain seedlings for an earlier and succesful transplant. Inoculation of
Lisianthus seeds inoculated with 106 cfu of A. brasilense showed the best
performance of plantlets.
Azospirillum is widely known for its ability to promote seedling
development, and it is postulated that the mechanisms involved in this
process are multiple yet to be fully understood (Bashan and De Bashan,
2010) Hernández et al. (1997). and Martín et al. (2004) reported that an
overdose of bacteria could cause an inhibitory effect in the plant, since in
sterile soil there are no other microorganisms that interact with Azo­
spirillum, allowing the production of hormones to accumulate (mainly
auxins, cytokinins and gibberellins), causing a depressive effect. Thus, a
lower dose of the rhizobacteria in sterile medium could have enhanced
its promoting capacity on plants, while the with 107 cfu did not generate
detrimental effects on the biometric parameters studied, but did not
generate differences with respect to treatment without either inoculate.
Cassán et al. (2009) showed that the Az39 strain of A. brasilense is
capable of improving the germination power of inoculated Glycine max
L. (Fabaceae) and Zea mays L. seeds, as well as the early development of

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M.P. Santos et al. Scientia Horticulturae 299 (2022) 111041

Table 1
Effect of Azospirillum brasilense Az39 bacterization on shoot and root parameters on in vitro germination of Eustoma grandiflorum after 60 days of culture.
Treatment Number of Leaf area Root Aerial Root biomass (mg)
leaves (cm2) length biomass (mg)
(cm)
Fresh weight Dry Fresh weight Dry weight
weight

Control 6.4 ± 0.4 b 0.3 ± 0.0 b 2.5± b 34.3 ± c 7.3 ± b 10.3 ± c 1.8 ± b
±0.2 1.7 0.9 2.3 1.4
Az39 106 cfu 10.2 ± 0.8 a 1.2 ± 0.1 a 8.2 ± a 988.9 ± a 38.8 ± a 605.2 ± a 46.0 ± a
0.2 15.3 2.3 24.0 4.0
7
Az39 10 cfu 8.1 ± 0.4 b 0.4 ± 0.0 b 6.6 ± b 201.4 ± b 12.7 ± b 65.4 ± b 2.7 ± b
0.2 4.4 1.1 3.8 1.1
Increases expressed as
percentages respect to
control
Control – – – – – – –
Az39 106 cfu 59 300 228 2783 431 5775 2455
Az39 107 cfu 27 33 164 487 74 535 50

Control: uninoculated seedlings; Az39 106 cfu: inoculation at seeding with 1.3 × 106 cfu of A. brasilense Az39; Az39 107 cfu: inoculation at seeding with 1.3 × 107 cfu of
A. brasilense Az39. Different letters indicate significant differences (p ≤ 0.05; Tukey’s test) between treatments for each parameter.

Fig. 3. Leaf transverse section of E. grandiflorum in vitro seedlings. (A), non-inoculated seedlings (control); (B), inoculation at seeding with A. brasilense Az39 1.3 ×
106 cfu of Az39; (C) inoculation at seeding with A. brasilense Az39 1.3 × 107 cfu. Scale bars: 50 µm.

Table 2
Morphometric analysis of in vitro E. grandiflorum leaves: control and Az39-inoculated treatments.
Thickness (µm) Cell size (µm)
Treatment Leaf Mesophyll Adaxial epidermis Abaxial epidermis

Control 120.2 ± 1.8 c 106.5 ± 1.6 c 8.7 ± 0.3 b 8.0 ± 0.5 b 8.4 ± 0.5 c
Az39 106 cfu 351.3 ± 6.3 a 311.8 ± 5.6 a 17.6 ± 0.4 a 14.4 ± 0.6 a 20.9 ± 0.9 a
7
Az39 10 cfu 245.8 ± 8.2 b 214.2 ± 7.1 b 19.8 ± 0.5 a 15.9 ± 0.9 a 10.7 ± 0.5 b
Increases expressed as percentages respect to control
Control – – – – –

Az39 106 cfu 192 193 102 80 149

Az39 107 cfu 104 101 128 99 27
Control: uninoculated seedlings; Az39 106 cfu: inoculation at seeding with 1.3 x 106cfu of A. brasilense Az39; Az39 107 cfu: inoculation at seeding with 1.3 x 107 cfu of A. brasilense

Fig. 4. Root transverse section of E. grandiflorum in vitro seedlings. (A), non-inoculated seedlings (control); (B), inoculation at seeding with A. brasilense Az39 1.3 ×
106 cfu of Az39; (C) inoculation at seeding with A. brasilense Az39 1.3 × 107 cfu. Scale bars: 50 µm.

4
M.P. Santos et al.
Table 3
Morphometric analysis of in vitro E. grandiflorum roots: control and Az39-inoculated treatments.
Diameter (µm) Thickness (µm)
Treatment Root Vascular bundle
Control 267.8 ± 10.1 c 73.1 ± 1.3
Az39 106 cfu 882.1 ± 15.1 a 185.6 ± 0.7
Az39 107 cfu 766.6 ± 17.1 b 72.9 ± 0.4
Control: uninoculated seedlings; Az39 106 cfu: inoculation at seeding with 1.3 × 106 cfu of A. brasilense Az39; Az39 107 cfu: inoculation at seeding with 1.3 × 107 cfu of
A. brasilense Az39. Different letters indicate significant differences (p ≤ 0.05; Tukey’s test) between treatments for each parameter.
Zarlavsky, G. E. (2014). Histología Vegetal: técnicas simples y complejas. Sociedad Argentina de Botánica: Buenos Aires.
seedlings. Effects of bacterization on germination may be due to the
major hormones synthesis that cause root initiation and cell elongation,
promote cell division and tissue expansion (Vessey, 2003; Pereg et al.,
2016). The germination percentage was low when compared to data
reported by Barbaro et al. (2009) in Lisianthus grown in a floating tray
system. They did not use any disinfection methodology so seed disin­
fection techniques used in our work may have affected seeds viability. It
would be necessary to determine other disinfection methodologies for
seeds of this species for future research.
The ability of A. brasilense to promote growth has been demonstrated
in numerous species. For example, significant increases in Photinia
(Photinia × fraseri Dress) root biomass have been described (Larraburu
et al., 2007) and in radical length of micropropagated shoots of apple,
Malus domestica (Borkh.) (Rosaceae) (Carletti et al., 2003) Verma et al.
(2011). also observed higher plant height and number of branches, in
addition to dry matter accumulation and higher number of flowers per
plant in Chrysanthemum morifolium Ramat (Asteraceae). In seedlings of
Petunia hybrida (cv Bravo White) foliar sprayed inoculants, presented
significant increases in height, number of shoots, leaf area and dry
weight of stem and roots. Positive effects were also observed at the
beginning of flowering (Nammidevi et al., 2008).
The increase on the number of adventitious roots presented by
inoculated treatments, fundamentally the treatment with 106 cfu of
A. brasilense is explained because one of the best described effects is the
impact on radical growth and development by Azospirillum sp. (Fukami
et al., 2018). It has been suggested that the increase in mineral ab­
sorption by plants is indirectly due to an increase in volume of the root
system, especially in the functionally active hair zone (Wang et al.,
2016). Anatomical studies provide information on the organization of
the plant’s organs allowing hypotheses to be made about the effect of
different treatments on plant development (Apóstolo et al., 2005;
Hazarika, 2006).
In anatomical study of leaf tissue, Larraburu & Llorente (2015)
demonstrated an increase in the structural size of the leaves of pink
lapacho H. impetiginosus (Bignoniaceae) in vitro inoculated with
A. brasilense. The increase in the thickness of the mesophyll and the
epidermis were in agreement with the increases in fresh and dry weight.
Furthermore, in photinia, inoculation with the Cd and Sp7 strains of
A. brasilense and Azotobacter chroococum induced an increase in the
thickness of the chlorenchyme, due to parenchymal cell proliferation in
palisade (Larraburu et al., 2010).
In the case of E. grandiflorum, it has been shown that inoculation with
Az39 (in both concentrations studied), promotes increases in the
thickness of the mesophyll and the epidermis due to the increase in cell
size of the chlorenchyma. The ability of PGPRs to produce phytohor­
mones and modify the metabolism of endogenous regulators (Bashan
and DeBashan, 2010), could be an explanation of these structural
changes provided in bacterial treatments compared to control. There­
fore, changes in the plant’s hormonal balance during interaction with
the bacteria would stimulate growth and plant cell division.
Rego et al. (2014) detected changes in the anatomical characteristics
of the roots of rice, Oryza sativa L. (Poaceae), when inoculated with
Azospirillum sp., such as the diameter of the vascular cylinder, the
thickness of the cortex, the rhizodermis and the cell size. Coinciding
Scientia Horticulturae 299 (2022) 111041
Crust Rhizodermis Cell size (µm)
b 75.1 ± 4.5 b 3.7 ± 0.1 c 11.3 ± 0.9 c
a 324.7 ± 10.4 a 7.5 ± 0.1 a 29.5 ± 1.5 a
b 256.7 ± 10.0 a 4.5 ± 0.2 b 18.4 ± 1.2 b
with these authors, significantly higher values were found in inoculated
treatments in Lisianthus compared to the control in all analyzed pa­
rameters, except for the diameter of the central cylinder, where the
significant differences only occurred in the treatment inoculated with
106 cfu. Lisianthus roots corresponding to bacterial treatments show
significant increases in the thickness of the cortex, the diameter of the
vascular cylinder and, consequently, in the total diameter of the root.
These results together with the increases in length and root biomass
reflect the potential of Az39 strain to promote root development.
The presence of conspicuous aerenchyma with large intercellular
spaces observed in the roots of the seedlings inoculated with the 107 cfu
treatment was evident. This could be explained as a process of
morphogenesis generated by the in vitro culture technique itself,
accentuated by a possible effect of the bacterial concentration that
would generate hypoxia, causing this anatomical modification that was
not seen in the other treatments. The profuse aerenchyma appears in
roots subjected to soils poor in oxygen (hypoxia) due to its high water
content. For example, aerenchyma development was observed in millet,
Panicum miliaceum. L (Poaceae) seedlings subjected to hypoxia (Dantas
et al., 2001).
The morpho anatomical changes generated by bacterization in
E. grandiflorum seedlings obtained in vitro with respect to the control,
could increase survival during the acclimatization phase. A greater
survival of micropropagated seedlings, also, was observed when inoc­
ulating with PGPRs other species cultivated in vitro, such as potato H.
impetiginosus (Bignoniacea) (Larraburu and Llorente, 2015), Photinia x
fraseri (Rosaceae) (Larraburu et al., 2007), jojoba Simmondsia chinensis L.
(Simmondsiaceae) (Larraburu et al., 2016a), tomato Lycopersicon escu­
lentum P. Mill. (Solanaceae) (Nowak and Shulaev, 2003), grapevine Vitis
vinífera L. (Vitaceae) (Barka et al., 2002), Alpinia purpurata K. Schum
(Zingiberaceae) (Ayora-Talavera et al., 2007) and Prunus sp. (Rosaceae)
(Russo et al., 2008).
Rosenblueth, 2006
Conclusions
Biofertilization of E. grandiflorum seeds increased in vitro germination
rate and produced precocity. Inoculation with 106 cfu of A. brasilense
Az39 promoted aerial and radical development evidenced by increases
in biometric parameters and anatomical modifications.
Therefore, the Az39 strain of A. brasilense, specially 106 cfu con­
centration, improved root development, generating homogeneous
seedlings, larger, healthy and vigorous. These features could be linked to
a better resistance to transplantation. The use of PGPR as A. brasilense
may be an useful tool in Lisianthus production, reducing times and costs.
Author contributions
MPS, SMC and EEL conceived and designed research and then
analyzed the obtained data. MPS, MEY and SJM conducted experiments.
SMC and EEL supervised the work. MPS, MEY, SMC y EEL wrote the
manuscript. All authors read and approved the manuscript.
5
M.P. Santos et al.
Credit author statement
María Paula Santos: Conceptualization, Methodology, Formal
analysis
Sergio Javier Martinez: Conceptualization, Methodology, Formal
analysis
Mauro Enrique Yarte: Investigation. Visualization. Writing- Original
draft preparation.
Susana Margarita Carletti: Supervision. Writing- Original draft
preparation.
Ezequiel Enrique Larraburu: Supervision. Visualization. Data cura­
tion. Writing- Reviewing and Editing. Project administration and
Funding acquisition.
Declaration of Competing Interest
Lisianthus Eustoma grandiflorum (Raf.) Schinners (Gentianaceae) is
an ornamental plant excellent for cut variety and it propagated from
seeds, which are very small and difficult to grow. The production of
seedlings using conventional systems is usually not efficient, obtaining
heterogeneous individuals with reduced aerial and root development.
For these reasons, it is necessary to find biotechnological tools such us
biofertilization to improve the propagation process. Our work explore
the effect of A. brasilense Az39 on E. grandiflorum and proposes bio­
fertilization as a strategy to improve the crops of this species.
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