The use of three fungal strains in producing of indole-3-acetic acid and gibberelllic acid

Abstract

In this work, the effective Indole-3-acetic acid (IAA) and Gibberellic acid (GA) producing conditions by Trichoderma harzianum UzCF-55, Penicillium canescens UzCF-54, Fusarium moniliforme UzGC-12 fungal strains have been investigated. It was found that the optimal pH of the nutrients for the strains was 5.5 for Mandel's and 6.8 for Czapek's mediums, with the incubation temperature intervals of 28-30°C for 10 days. Quantitative analysis of the IAA synthesized by T. harzianum UzCF-55 revealed that the highest amount of the acid was 1,16 mg/mL and 0.74 mg/mL, respectively on the sixth day of the exponential phase of micromycetes growth, while it was 0.318 mg/ml and 0.17 mg/mL for the GA after ninth day. The amounts of IAA and GA synthesized by P. canescens UzCF-54 were 0.98 mg/mL and 0.38 mg/mL in the similar days of incubation, also showing greater amounts than the control strain. F. moniliforme UzGC-12 strain synthesized 0.63 mg/mL IAA and 0.39 mg/mL GA, respectively. High performance liquid chromatography-mass spectrometry (HPLC-MS) analyses of the micromycetes revealed the mass fractions of 174.00 m/z corresponding to the molecular mass of IAA, 363.00 m/z of GA7 and 361.00 m/z of GA3, indicating gibberellins syntheses by strains.

Full text

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32
Original Research Article
Plant Cell Biotechnology and Molecular Biology 21(35&36):32-43; 2020 ISSN: 0972-2025
THE USE OF THREE FUNGAL STRAINS IN PRODUCING
OF INDOLE-3-ACETIC ACID AND GIBBERELLLIC ACID
BAKHORA TURAEVA
*
, AZAM SOLIEV, FARKHOD ESHBOEV,
LUKHMON KAMOLOV, NODIRA AZIMOVA, HUSNIDDIN KARIMOV,
NIGORA ZUKHRITDINOVA AND KHURSHIDA KHAMIDOVA
Institute of the Microbiology, Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan
[BT, NA, HK, NZ, KK].
Republican Scientific and Practical Center of Sports Medicine, Tashkent, Uzbekistan [AS].
Institute of the Chemistry of Plant Substances, Academy of Sciences of the Republic of Uzbekistan,
Tashkent, Uzbekistan [FE].
Karshi State University, Kashkadarya region, Uzbekistan [LK].
[
*
For Correspondence: E-mail:
turaevabakhora@mail.ru, jaloliddinshavkiev1992@gmail.com]
Article Information
Editor(s):
(1)
Dr. Ravi Kant Chaturvedi, Chinese Academy of Sciences, P. R. China.
Reviewers:
(1)
Javad Sahandi, Ocean University of China, China.
(2)
Muhammad Adeel Manzoor, University of Veterinary and Animal Sciences, Pakistan.
Received: 15 June 2020
Accepted: 20 August 2020
Published: 26 August 2020
_______________________________________________________________________________________________
ABSTRACT
In this work, the effective Indole-3-acetic acid (IAA) and Gibberellic acid (GA) producing
conditions by Trichoderma harzianum UzCF-55, Penicillium canescens UzCF-54, Fusarium
moniliforme UzGC-12 fungal strains have been investigated. It was found that the optimal pH of the
nutrients for the strains was 5.5 for Mandel’s and 6.8 for Czapek's mediums, with the incubation
temperature intervals of 28-30°C for 10 days. Quantitative analysis of the IAA synthesized by T.
harzianum UzCF-55 revealed that the highest amount of the acid was 1,16 mg/mL and 0.74 mg/mL,
respectively on the sixth day of the exponential phase of micromycetes growth, while it was 0.318
mg/ml and 0.17 mg/mL for the GA after ninth day. The amounts of IAA and GA synthesized by P.
canescens UzCF-54 were 0.98 mg/mL and 0.38 mg/mL in the similar days of incubation, also
showing greater amounts than the control strain. F. moniliforme UzGC-12 strain synthesized 0.63
mg/mL IAA and 0.39 mg/mL GA, respectively. High performance liquid chromatography-mass
spectrometry (HPLC-MS) analyses of the micromycetes revealed the mass fractions of 174.00 m/z
corresponding to the molecular mass of IAA, 363.00 m/z of GA7 and 361.00 m/z of GA3, indicating
gibberellins syntheses by strains.
Keywords: T. harzianum; P. canescens; F. moniliforme; fungi; gibberellic acid; indole-3-acetic acid.
INTRODUCTION
Ever-increasing population of the earth needs a
sustainable agricultural industry that requires huge
amounts of chemical fertilizers to be used in this
sector. However, such amount of fertilizers may
cause huge damages to the environment and
human health [1]. The application of biofertilizers,

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33
in the contrary, have many advantages as they are
environmentally friendly, enhancing crop
productivity, and economically valuable [2].
Phytohormones play an important role in
agriculture and medicine, which is reflected in an
industrial production of phytohormones,
especially using fungal cultures [3].
Phytohormones are divided into five main groups:
auxins, gibberellins, cytokines, ethylene, and
abscisic acid [4,5]. They are able to modify the
physiological functions of plants to accelerate
their growth by intensive cell division in callus
tissue, promote phloem development, enhance
lateral root development, stimulate plant growth,
prevent leaf aging by slowing down the
breakdown of chlorophyll pigments in plants, and
improve metabolism even at low concentrations.
The phytohormones indoleacetic acid, gibberellic
acid and ethylene are synthesized not only by
plants, but also by many soil and rhizosphere
microorganisms [6,7,8,9]. For example, when
seeds were inoculated with a culture fluid of the
fungal strains of Trichoderma harzianum and
Penicillium canescens root development and
nutrient uptake were increased by 76 and 61%,
respectively, with the percentages of IAA and GA
that increased by 49 and 71% in plant leaves and
40 and 143% in roots, accordingly [10].
IAAs are responsible for the division, extension,
and differentiation of plant cells and tissues.
Phytohormones of this group stimulate seed and
tuber germination; increase the rate of xylem and
root formation; control processes of vegetative
growth, tropism, fluorescence, and fructification
of plants, affecting photosynthesis, pigment
formation, biosynthesis of various metabolites,
and resistance to stress factors. However, at high
concentrations, they can inhibit plant growth [4,5].
GAs are diterpenoid plant hormones, first isolated
from the phytopathogenic fungus Gibberella
fujikoroi in the 1920s [11,12,13,14]. Gibberellins,
classified with diterpenes, consist of isoprene
residues that usually form four rings. GAs GA3,
GA7, GA1, and GA4 are the best studied
phytohormones of this group. They exhibit
maximum biological activity and are the most
widespread in nature. GAs participate in every
aspect of plant growth and development including
seed germination, stem extension, flowering,
aging and stimulate the formation of hydrolytic
enzymes in germinating cereal grain [11,7,4,5].
The formation of IAA and GA are widespread
among fungi of the genera Fusarium, Rhizoctonia,
Rhizopus, Absidia, Aspergillus, Penicillium,
Monilia, Phoma, Pythium, Trichoderma,
Verticillum, Schizophillum, and Actinomucor
[11,7.12,13,]. For instance, active GAs were
studied in 28 strains of Fusarium moniliform
fungus, but in 4 strains was observed a high level
of activity and the maximum amount of
synthesized GA was 2.40 g/L [15]. Therefore,
fungi are greatly important on the growth and
development of plants. Because, fungi being
remarkable organisms that readily produce a wide
range of secondary metabolites such as
phytohormones and mycotoxins [16]. However,
the influence of various carbon sources, pH of
nutrient medium (5.7 and 8) and temperature (23,
25, 30, 37°C) on the formation of active GA and
IAA by the fungal strains have not been well
studied yet. It is reported that high amount of GA
(5.8 g/L and 105.0 mg/g) were obtained when F.
moniliforme fungal strain was grown for 4 days at
30°C in a solid and liquid Czapek's nutrient
medium enriched with Jatropha curcas L. plant
residue as a carbon source. The use of plant waste
materials as a source of carbon increases the
amount of GA production up to 2.5 times [17,18].
The aim of this study was to investigate the impact
of different carbon sources, pH, and temperature
on the formation of GA and IAA by the
rhizosphere fungi strains of T. harzianum UzCF -
55, P. canescens UzCF-54 and F. moniliforme
UzGC-12 with the ultimate goal of creation of a
plant growth promoting fungal (PGPF) fertilizer.
MATERIALS AND METHODS
Extraction and Identifications of GA and IAA
from Culture Fluids of Micromycetes T.
harzianum-55, P. canescens-54 and F.
moniliforme-12
The studied fungal strains were isolated from the
rhizosphere of agricultural plant (cotton and corn)
roots in laboratory (Bioconversion of plant raw
materials) and patented [19,20,21].
The selected strains were inoculated in above
mentioned nutrient mediums and incubated for ten

Turaeva et al.
34
days by shaking at 220 rpm (SK-L330-Pro, China)
with the temperature set at 28°C. After incubation,
the culture broth was filtered using sterilized 0.22
μm bacteriological filter. The pH of the isolated
culture fluid was adjusted to 2.0–2.2 using 2 M
HCl and extracted 3 times with ethyl acetate
(EtOAc). The extracted IAA and GAs from T.
harzianum-55, P. canescens-54 and F.
moniliforme-12 were then subjected to TLC,
HPLC and HPLC-MS analyses.
TLC analysis of GA and IAA. TLC identification
of GA and IAA were performed on silica gel
plates (Silica gel on TLC AL foils L × W 10 cm ×
20 cm Gf254 (Germany)) in an n-hexane-ethyl
acetate – 95:5 (v/v) system. Pure GA and IAA
(Sigma Aldrich) were used as standards.
HPLC analyses of GA and IAA. To perform
HPLC analyses of the samples the extracted
culture fluids of the micromycetes (sixth and ninth
days) were filtered through a 0.2 μm filter. HPLC
analyses of GA and IAA from the filtered culture
fluids were performed on an Agilent series 1200
(USA) chromatographic system, consisting of a
3085PU micro pump, a 3080DG degasser, a
3067CO column furnace, a 3070UV UV detector,
and a 3080MX mixer. Substances were separated
using a C18 modified Chromabond column (250.0
mm × 2.1 mm) filled with a particle size of 5 μm.
HPLC analysis of GA was carried out using the
condition of UV detector set at 204 nm
wavelength and at 30°C. The mobile phase used
was water/acetonitrile – 80:20 (v/v) at a flow rate
of 1.5 ml/min. The sample injection volume was
10 μl. The retention time of sample peaks was
compared with those of pure GA (Sigma Aldrich)
standards [22].
HPLC analysis of IAA was performed at a UV
detector of 280 nm and at 30°C in the mobile
phase of water/acetonitrile – 85:15 (v/v) with a
flow rate of 1.5 ml/min. The sample injection
volume was 10 μl. The retention time of sample
peaks was compared with those of authentic IAA
(Sigma Aldrich) standards [22].
Study of Formulation of GA and IAA
Determination of gibberellin acid content. The GA
concentration was determined using a modified
method described by Hasan [23]. The pH of
filtrates was adjusted to 2.7 with 1 M HCl and
extracted three times with equal volumes of ethyl
acetate. The ethyl acetate fractions were collected
and vacuum evaporated at 20°C. The dry pellet
was re-suspended in ethanol/H2SO4 (9:1 v/v)
mixture. The absorbance was measured at λ = 254
nm (Spekol 1300) and the GA concentration was
calculated from the regression equation of a
standard curve of pure gibberellic acid (Sigma-
Aldrich) and expressed as µg/mL.
Determination of indoleacetic acid. The IAA
concentration in the culture supernatant was
determined according to the method of Jaroszuk
[7]. The filtered 1.0 mL of the cultura fluid and
1.0 mL of Salkowski’s reagent (50 mL solution of
35% H2SO4 and 1 mL of 0.5 M FeCl3 mixture)
were mixed intensively then the reaction mixture
stayed for 30 minutes. The samples turned to
reddish-pink and the concentration of IAA was
determined by measuring the optical density of the
samples through a green light filter at a
wavelength of 535.0 nm by spectrophotometer
(Spekol 1300). The indoleacetic acid
concentration was calculated using a calibrated
graph of the amount of pure indoleacetic acid
(Sigma Aldrich). It was expressed as μg/mL.
Study of IAA and GA Production in Different
Growth Conditions
The suspensions of 106 spore/mL concentrations
of T. harzianum UzCF-55, P. canescens UzCF-54
and F. moniliforme UzGC-12 strains grown for 6
days in Mandel’s and Czapek's nutrient mediums
(in solution) were used as planting material. In this
study, Mandel’s and Czapek's mineral nutrient
mediums
were
used
for
the
culture of T. harzianum
UzCF-55, P. canescens UzCF-54 and F.
moniliforme UzGC-12 strains in order to study the
GA and IAA production capacity of these strains.
Mandel’s mineral nutrient medium for
Micromycetes included (in g/l): KH
2
PO
4
– 2.0;
(NH
4
)
2
HPO
4
– 1.4; MgSO
4
– 0.5; CaCl
2
– 0.3;
sucrose – 2.0; microelements – 1 ml; (500 mg
FeSO
4
; 156 mg MnSO
4
·4Н
2
О; 167 mg ZnCl
2
; 200
mg CoCl
2
; 19 % HCl in 100 ml distilled water)
[24]. Czapek's medium consisted of (in g/l):
KH
2
PO
4
– 1.0; MgSO
4
- 0.5; NaNO
3
– 3.0; KCl –
0.5; FeSO
4
- granules; sucrose - 2; microelements
- 1 ml; (mixture of 500 mg FeSO
4
; 156 mg

Turaeva et al.
35
MnSO
4
, 4N
2
O; 167 mg ZnCl
2
; 200 mg CoCl
2
; 1
ml 19% HCl in 100 ml distilled water) [3].
Additionally, molasses, 2% corn extract, 2%
powdered cotton stalks and cotton leaves were
added as a carbon source for both nutrient
mediums. Additionally, the effects of initial pH
value and the temperature of nutrient media on the
formation of GAs and IAA by these fungal strains
were also studied. The pH ranges from 4.0 to 7.5
(4.0; 5.5; 5.0; 5.5; 6.0; 6.5; 6.8; 7.0; 7.5;) and the
temperature from 20°C to 45°C (20°C; 25°C;
28°C; 30°C; 35°C; 40°C;) were tested for 10 days
in a shaker at 200–220 rpm/min in the growth of
the strains.
HPLC-MS Analysis of GA and IAA
To perform HPLC-MS analyses of the GA and
IAA samples the extracted culture fluids of the
fungal strains of T. harzianum-55, P. canescens-
54 and F. moniliforme-12 were filtered through a
0.22 μm bacteriological filter. Analyses were
performed on an Ultimate 3000 Quaternary
Standard UHPLC connected to TSQ Quantum
Access Max triple quadrupole mass-spectrometer
(UHPLC-МS/МS) (Thermo Fisher Scientific,
USA). Samples were injected using an MS auto
sampler unit. The UHPLC column used was a
HyperSil Super Gold C18 (100 mm×2.1 mm, 1.9
µm) (Thermo Fisher Scientific, USA). Mobile
phase used was acetonitrile – water (added either
0.1% formic acid for positive ion monitoring or
0.1% ammonium acetate for negative ion
monitoring) in a gradient elution mode by
increasing the amount of organic solvent up to
80% at a flow rate of 0.2 mL/min. The column
temperature was set at 254ºC. Sample injection
volume was 1 µL. The quadrupole temperature
was set at 270ºC. The MS was operated in full
scan mode using either positive or negative ion
monitoring. MS conditions were set as following:
Instrumentation: Thermo Fisher Scientific TSQ
Quantum Access Max triple quadrupole mass-
spectrometer. Ionization condition – HESI; Spray
voltage – 2500 V for negative and 4000 V for
positive ion monitoring; Vaporizer temperature
(ºC) – 300; Sheath gas pressure (psi) – 35; Aux
gas pressure (Arb) – 10; Capillary temperature
(ºC) – 350; Tube Lens Offset – 80; Collision
pressure (mTorr) – 1.5; Collision energy (eV) –
10.
Statistical Analysis
The experiments were carried out in three
independent replicates and the results were
expressed as mean standard deviations (SD). The
SD values (represented as deviation bars) were
determined using Microsoft Excel 2016
(Microsoft Corp., Redmond, Washington, DC,
USA). Data analysis was performed using
StatView (SAS Institute Inc., Cary, NC, USA)
with one-way ANOVA followed by a Tukey’s
post hoc test where applicable, with the
significance evaluated at p < 0.05.
RESULTS AND DISCUSSION
Identification of IAA and GA in Culture Fluids
of the Fungi
Identification methods such as TLC and HPLC are
important in the study of secondary metabolites
synthesized by microorganisms [25]. Because
these methods of identification give opportunities
to determine whether those substances being
analyzed produced or not by the selected
microorganisms. Therefore, TLC and HPLC
identification were performed in this study in
order to realize the formation of GA and IAA by
the selected fungal strains. According to the
results of the TLC and HPLC analysis of the
culture fluid of micromycetes, it was found that
the GA and IAA were produced by all three fungal
strains studied in this work. Their retention factor
(Rf) and retention time (Rt) points were shown in
Table 1.
Table 1. Rf and Rt points of GA/IAA within culture fluids of selected fungal strains
Samples
(culture fluids of the strains)
Rf points Rt points
IAA(mg/mL) GA(mg/mL) IAA(mg/mL) GA(mg/mL)
T.harzianum UzCF-55 0.069 0.098 1.316 1.910
F.moniliforme UzGC-12 0.068 0.096 1.302 1.889
P.сanescens UzCF-54 0.095 0.127 1.340 1.944
Standards 0.074 0.105 1.308 1.910

Turaeva et al.
36
As can be seen from Table 1, the GA and IAA
produced by T. harzianum UzCF-55, P. canescens
UzCF-54, and F. moniliforme UzGC-12 were
identical compared to standards. The optimum
cultivation conditions of the fungal strains such as
pH, temperature, and carbon sources have also
been investigated to find the highest amount of
phytohormones by the fungi.
Influence of Different Carbon Sources, pH, and
Temperature on the Formation of GA and IAA
by Strains of T. harzianum UzCF-55, P.
сanescens UzCF-54 and F. moniliforme UzGC-
12
The incubation conditions such as pH, temperature
as well as carbon source and nitrogen are greatly
affect for the production of GA and IAA by the
fungi [26]. In this work, the influence of the
nutrient media was also studied in order to
increase the production of GA and IAA by T.
harzianum UzCF -55, P. сanescens UzCF -54, F.
moniliforme UzGC -12 strains by addition of 2%
sucrose and other carbon sources such as
molasses, cotton stalk powder, crushed cotton leaf,
corn extract. The production of GA and IAA was
studied in dynamics. As a result, it was found that
all three strains produced the highest amounts of
GA and IAA in nutrient media enriched with corn
extract. Additionally, the P. canescens UzCF-54
synthesized higher amounts of IAA and GA
compared to T. harzianum UzCF -55, and F.
moniliforme UzGC -12 when they grew in nutrient
media enriched with corn extract and molasses
where the amounts of IAA and GA reached up to
0.372 mg/mL, 0.310 mg/mL, 0.349 mg/mL and
0.294 mg/mL respectively (Fig. 1).
The optimum pH conditions were found to be 5.5,
6.5 and 6.8 for the production of phytohormones
by T. harzianum UzCF-55, P. сanescens UzCF-54
and F. moniliforme UzGС-12, respectively
(Fig. 2). The highest value of IAA and GA
synthesized by these strains of fungi had
occurred in the temperature range of 28–30°C
(Fig. 3).
After optimization of the cultivation conditions of
fungi for the formulation of phytohormones, the
micromycetes were grown in those conditions and
studied their phytohormones producing capacity
(Table 2).
A
0
0.2
0.4
0.6
0.8
1
1.2
control
cotton leaves
powdered
cotton stalks
molasses
corn extract
The amount of IAA (mg/ml)
T. harzianum UzCF-55
P. сanescens UzCF-54
F. moniliforme UzGC-12

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37
B
Fig. 1. The amount of IAA (A) and GA (B) produced by the strains of T. harzianum UzCF-55, P.
сanescens UzCF-54 and F. moniliforme UzGC-12 in media enriched with different carbon sources
A
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
control cotton leaves powdered cotton
stalks
molasses corn extract
The amount of GA (mg/ml)
T. harzianum UzCF-55 P. сanescens UzCF-54 F. moniliforme UzGC-12
0
0.1
0.2
0.3
0.4
0.5
0.6
4 4.5 5 5.5 6 6.5 6.8 7 7.5
The amount of GA (mg/ml)
pH value
P. сanescens UzCF-54 F. moniliforme UzGC-12 T. harzianum UzCF-55

Turaeva et al.
38
B
Fig. 2. Effect of the pH conditions of the nutrient medium on the production of GA (A) and IAA (B)
by the studied fungal strains
A
0
0.2
0.4
0.6
0.8
1
1.2
4
4.5
5
5.5
6
6.5
6.8
7
7.5
The amount of IAA (mg/ml)
pH value
F. moniliforme UzGC-12 P. сanescens UzCF-54 T. harzianum UzCF-55
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.5
20
25
28
30
35
40
45
The amount of GA (mg/ml)
Temperature, °C
P. сanescens UzCF-54
F. moniliforme UzGC-12
T. harzianum UzCF-55

Turaeva et al.
39
B
Fig. 3. Effect of the temperature on the producing of GA (A) and IAA (B) by the studied fungal
strains
Table 2. The amount of phytohormones synthesized by selected fungal strains
The strains of selected micromycetes GA
(mg/mL)
IAA
(mg/mL)
T.harzianum UzCF-55 0.386 ±0.004 0.635 ± 0.001
P.canescens UzCF-54 0.325 ± 0.001 1.167 ± 0.002
F.moniliforme UzGC-12 0.396 ± 0.002 0.984 ± 0.001
The HPLC-MS analyses of IAA and GA in culture
fluids of the fungal strains were conducted to
determine phytohormone-producing capacities of
fungi. The obtained results of the full mass spectra
of the culture fluids revealed the mass fragments
of m/z 174, m/z 361, and m/z 363 that correspond
to IAA, GA
3
and GA
7
, respectively (Fig. 4).
The obtained results indicated that the
phytohormones were produced intensively by the
selected fungal strains even when they had grown
together. These findings give an opportunity to
obtain the plant growth promoting fungal
fertilizers based on these fungi.
The biofertilizers are gaining significance
influence in sustainable agriculture as a means of
enhancing crop productivity, economically viable
manner, and reducing the polluting effect of
synthetic fertilizers [27,28]. Thus, in the present
study, phytohormones producing capacities of
selected fungal strains were studied to develop a
plant growth promoting fungal biofertilizers based
on these fungal associations.
0
0.2
0.4
0.6
0.8
1
1.2
20 25 28 30 35 40 45
The amount of IAA (mg/ml)
Temperature, °C
F. moniliforme UzGC-12 P. сanescens UzCF-54 T. harzianum UzCF-55

Turaeva et al.
40
Fig. 4. Representative mass spectrum of the micromycetes culture fluid
There are numerous reports on the identification
of IAA and GA by TLC and HPLC methods [29].
In a study by Tosapon [30], n-hexane:ethyl
acetate:isopropanol: acetic acid (40:20:5:1) system
was used as a mobile phase for the determination
of IAA by the TLC method from C. fructicola
CMU-A109 where its Rf value was 0.68. In our
study, n-hexane and ethyl acetate were also used
as a mobile phase for the determination of IAA
from the culture media of T. harzianum UzCF-55,
P. сanescens UzCF-54 and F. moniliforme UzGС-
12 where the Rf points of IAA were 0.069, 0.095
and 0.068, respectively. According to Kirti [31],
the Rf values of GAs were from 0.20 to 0.80.
However, in our work the Rf values of GA in
culture extracts of T. harzianum UzCF-55, P.
сanescens UzCF-54 and F. moniliforme UzGС-12
were 0.098, 0.096 and 0.127, respectively. The
reason for the difference in Rf values are the
different experimental conditions performed by
the two investigations. However, the Rf points of
phytohormones were appropriate to the Rf points
of the standards. Additionally, different HPLC
conditions of IAA and GA were created in
previous studies by the authors and they
established different Rt values [32,31,29]. In our
study, the Rt values of IAA and GA were also
consistent with the standards indicating that IAA
and GA were synthesized by selected fungal
strains (Tab.1). Most of the previous researchers
focused on the impact of carbon sources, pH, and
temperature in order to increase the productions of

Turaeva et al.
41
IAA and GA by the rhizosphere microorganisms,
because of their significant contributions,
particularly to the maintenance of enhancing crop
yields [18,26]. In a study by Patricia [18] banana
peel was used as a carbon source for the
production of GA by Fusarium moniliforme and
Aspergillus niger in the optimized fermentation
conditions: pH 5.5; inoculum temperature 25 ±
2oC; fermentation time 7 days. According to Işil
et al., [31], the optimum conditions for the
production of indole-3-acetic acid by the
Aspergillus niger were 6 days of incubation at
25°C with pH 6.0 and for gibberellic acid
production they were 12 days of incubation at
30°C with pH 5.0. However, in the present study,
the optimum carbon source for the production of
GA and IAA by T. harzianum UzCF -55, P.
сanescens UzCF -54, F. moniliforme UzGС -12
was the corn extract. The molasses has also
shown good results; therefore, it can also be used
as a carbon source in biotechnological production
of IAA and GA because the price of molasses is
cheaper than corn extract. The optimum pH points
were determined to be 5.5, 6.5 and 6.8 at the
temperature ranges of 28–30°C for the
formulation of a higher amount of the
phytohormones by T. harzianum UzCF-55, P.
сanescens UzCF-54 and F. moniliforme UzGС -
12, respectively. These conditions were almost
similar with the literature data for the production
of maximum amounts of IAA and GA that can be
synthesized by these strains.
CONCLUSION
Current study deals with the production of IAA
and GA by new fungal strains of T. harzianum
UzCF-55, P. сanescens UzCF-54 and F.
moniliforme UzGС -12. The optimum carbon
source, pH value, and temperature has been
investigated for the production of higher amounts
of IAA and GA by these fungal strains. It was
found that the highest amounts of IAA (1.167
mg/ml) and GA (0.396 mg/ml) were synthesized
by P. canescens UzCF-54 and F. moniliforme
UzGC-12 respectively when the strains were
grown in the optimized cultivation conditions.
HPLC-MS analyses have revealed the presence of
IAA, GA
3
, and GA
7
when these strains of
micromycetes were cultivated together. The
obtained results from this study give an
opportunity of the studied fungal strains to be used
for the biotechnological production of IAA and
GA.
COMPETING INTERESTS
Authors have declared that no competing interests
exist.
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