© The Authors, 2024, Published by the Universidad del Zulia*Corresponding author: rosa.arias@inecol.mx
Keywords:
Phosphate solubilizing fungi (PSF)
Coee plants
Biofertilization
Biosolubilization of phosphate by strains of Trichoderma in vitro and in greenhouse in three
varieties of Coea arabica
Biosolubilización de fosfatos por cepas de Trichoderma in vitro y en invernadero sobre tres variedades
de Coea arabica
Biosolubilização de fosfatos por cepas de Trichoderma in vitro e em invernada sobre três variedades
de Coea arabica
Rosa María Arias Mota
1
*
Alberto Donaldo Torres Salas
1
Yamel del Carmen Perea Rojas
2
Yadeneyro de la Cruz Elizondo
3
Rev. Fac. Agron. (LUZ). 2024, 41(4): e244241
ISSN 2477-9407
DOI: https://doi.org/10.47280/RevFacAgron(LUZ).v41.n4.10
Crop production
Associate editor: Dra. Lilia Urdaneta
University of Zulia, Faculty of Agronomy
Bolivarian Republic of Venezuela
1
Instituto de Ecología A.C. km 2.5 antigua carretera a
Coatepec 361 Congregación el Haya, CP 91073, Xalapa,
Veracruz, México.
2
Doctorado en Micología Aplicada, Centro de Investigación
en Micología Aplicada, Universidad Veracruzana Médicos
No. 5, U. H. del Bosque, CP 91010, Xalapa, Veracruz,
México.
3
Facultad de Biología. Universidad Veracruzana, circuito
Gonzalo Aguirre Beltrán s/n. Zona Universitaria, CP 91000,
Xalapa, Veracruz, México.
Received: 12-09-2024
Accepted: 05-11-2024
Published: 18-11-2024
Abstract
Coee soils have a low availability of phosphorus, the use
of phosphate-solubilizing Trichoderma strains is a promising
sustainable strategy for the management of phosphorus deciencies.
In this study, we evaluated 10 strains of Trichoderma from the
andosol soil of coee plantations in Mexico and their capacity for
phosphate solubilization in vitro, and their impact on the growth
of coee seedlings of three varieties (Anacafé, Costa Rica and
Marsellesa) were evaluated. The tested microorganisms showed
high phosphorus solubilization, the phosphorus solubilization
ranged between 2.41 and 7.40 mg.mL
-1
. The maximum phosphate
solubilizing activity was observed using two strains of Trichoderma
harzianum 75.73 (Th53) and 74.62 mg.mL
-1
(Th48) for calcium
phosphate (Ca
2
PO
4
) and three strains of T. asperellum 22.99 (Th57),
22.90 (Th49) and 21.55 mg.mL
-1
(Th40) for aluminum phosphate
(AlPO
4
). In both calcium (Ca
2
PO
4
) and aluminum phosphate
(AlPO
4
), a decrease in the pH of the medium was detected, from
4.81 to 3.73 and from 3.38 to 2.75, respectively. In the Anacafé
variety, the application of two strains of T. harzianum (Th48 and
Th53) favored greater availability of phosphorus in the substrate,
while in the Costa Rica and Marsellesa varieties the available
phosphorus of the substrate was greater with T. harzianum (Th48).
Inoculation with these Trichoderma strains is potentially important
for the solubilization of insoluble phosphorus and the development
of coee plants.
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Rev. Fac. Agron. (LUZ). 2024, 41(4): e244141 October-December. ISSN 2477-9407.
2-7 |
Resumen
Los suelos cafetaleros presentan una baja disponibilidad de
fósforo, el uso de cepas de Trichoderma fosfato solubilizadoras es una
estrategia sostenible prometedora para el manejo de las deciencias
de fósforo. En este estudio, se evaluaron 10 cepas de Trichoderma
provenientes de suelos andosoles de cafetales en México para
determinar su capacidad de solubilización de fosfato in vitro, y su
impacto en el crecimiento de plántulas de café de tres variedades
(Anacafé, Costa Rica y Marsellesa). Los microorganismos probados
mostraron alta solubilización de fósforo, la solubilización de fósforo
osciló entre 2,41 y 7,40 mg.mL
-1
. La máxima actividad solubilizadora
de fosfato se observó utilizando dos cepas de Trichoderma harzianum
75,73 (Th53) y 74,62 mg.mL
-1
(Th48) para fosfato de calcio (Ca
2
PO
4
)
y tres cepas de T. asperellum 22,99 (Th57), 22,90 (Th49) y 21,55
mg.mL
-1
(Th40) para fosfato de aluminio (AlPO
4
). Tanto en fosfato de
calcio (Ca
2
PO
4
) como de aluminio (AlPO
4
) se detectó una disminución
del pH del medio, de 4,81 a 3,73 y de 3,38 a 2,75, respectivamente.
En la variedad Anacafé, la aplicación de dos cepas de T. harzianum
(Th48 y Th53) favorecieron una mayor disponibilidad de fósforo en
el sustrato, mientras que en las variedades Costa Rica y Marsellesa el
fósforo disponible del sustrato fue mayor con T. harzianum (Th48).
La inoculación con estas cepas de Trichoderma es potencialmente
importante para la solubilización del fósforo insoluble y el desarrollo
de las plantas de café.
Palabras clave: hongos solubilizadores de fosfato (PSF), plantas de
café, biofertilización.
Resumo
Os solos cafeeiros apresentam baixa disponibilidade de fósforo,
o uso de cepas de Trichoderma solubilizadoras de fosfato é uma
estratégia sustentável promissora para o manejo das deciências de
fósforo. Neste estudo, foram avaliados cepas 10 Trichoderma do
solos andossolos de plantações de café no México e sua capacidade
de solubilização de fosfato in vitro, e seu impacto no crescimento de
mudas de café de três variedades (Anacafé, Costa Rica e Marsellesa).
Os microrganismos testados apresentaram solubilização de fósforo
variou entre 2,41 e 7,40 mg.mL
-1
. A máxima atividade solubilizadora
de fosfato foi observada utilizando duas cepas de Trichoderma
harzianum 75,73 (Th53) e 74,62 mg.mL
-1
(Th48) para fosfato de
cálcio (Ca
2
PO
4
) e três cepas de T. asperellum 22,99 (Th57), 22,90
(Th49) e 21,55 mg.mL
-1
(Th40) para fosfato de alumínio (AlPO
4
).
Tanto no fosfato de cálcio (Ca
2
PO
4
) quanto no fosfato de alumínio
(AlPO
4
) foi detectada diminuição do pH do meio, de 4,81 para 3,73 e
de 3,38 para 2,75, respectivamente. Na variedade Anacafé, a aplicação
de duas cepas de T. harzianum (Th48 e Th53) favoreceu maior
disponibilidade de fósforo no substrato, enquanto nas variedades
Costa Rica e Marseillaise o fósforo disponível do substrato foi
maior com T. harzianum (Th48 ). A inoculação com estas cepas de
Trichoderma é potencialmente importante para a solubilização do
fósforo insolúvel e o desenvolvimento dos cafeeiros.
Palavras-chave: fungos solubilizadores de fosfato (PSF), café,
Biofertilização.
Introduction
Numerous studies have classied species of the genus Trichoderma
as benecial microorganisms for plants because they exhibit
antagonistic activity against phytopathogenic fungi, promote plant
growth, improve crop yield, promotes abiotic stress tolerance and
increases nutrient utilization (Zin and Badaluddin, 2020). In recent
years, phosphorus-solubilizing fungi (PSF) have been considered as
an alternative phosphorus fertilizer that can promote plant growth
(Yang et al., 2022). In Mexico, one of the main problems in coee
production is the low availability of phosphorus in the soil, as this
crop is often grown on volcanic soils characterized by acidic pH and
low availability of essential macronutrients, including phosphorus
(Geissert and Ibanez, 2008). The average phosphorus content in the
coee soils of the region is 1.75 mg.kg
-1
; according to Sadeghian
et al. (2019) is below the optimal level for coee cultivation (10-
30 mg.kg
-1
). Generally, soil phosphorus deciency can be solved by
adding phosphorus fertilizers; however, these lack eciencies due
to the rapid conversion of available phosphorus to insoluble forms.
Therefore, it is very important to study the phosphorus solubilizing
potential of Trichoderma species, because of their activity they can
act as biofertilizers. The objectives of this research were to estimate
the in vitro phosphorus solubilization capacity of ten Trichoderma
strains from the rhizosphere of coee trees and to evaluate their
eect on phosphorus availability and coee growth under greenhouse
conditions.
Materials and methods
Fungal material
Trichoderma strains were isolated from rhizospheric soil samples
from coee plantations in the center of the state of Veracruz (Arias et
al., 2022) using the soil particle washing technique. The strains were
reactivated in potato dextrose agar (PDA) culture medium and kept in
incubation (Thermo Scientic® 150) for 10 days at 25 °C in the dark.
Identication of the Trichoderma isolates
For molecular identication, mycelial DNA was extracted from
pure cultures and recover by scraping with a sterile needle. Genomic
DNA extraction was performed using the Wizard® Genomic DNA
Purication Kit from Promega (Madison, USA) according to the
manufacturer’s instructions. The nuclear rDNA ITS region (ITS1-
5.8S-ITS2) was amplied using ITS5/ITS4 primers (White et al.,
1990). Amplication was performed by PCR (polymerase chain
reaction) using MyTaq DNA polymerase (Meridian Bioscience).
For sequencing, PCR products were dispatched to Macrogen, Inc
(Seoul, Korea). The genetic anity of the obtained sequences was
determined using the BLASTn algorithm of the National Center for
Biotechnology Information GenBank (NCBI, 2024). Furthermore,
Trichoderma strains are identied according to the method of Gams
and Bisset (2002) by the morphological characteristics (mycelial
structure, concentric ring formation and conidial pigmentation of the
colonies) and the microscopic (shape and arrangement of conidia,
phialides and conidiophores).
Study quantitative of the ability of Trichoderma strains to
solubilize calcium phosphate and aluminum phosphate
For this measurement, Pikovskaya (1948) liquid culture medium
was used with tricalcium phosphate (Ca
3
(PO
4
)
2
) (0.5 g.L
-1
) or
aluminum phosphate (AlPO
4
) (0.5 g.L
-1
) as a source of insoluble
phosphorus. Each Trichoderma strain was inoculated with four discs
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Arias et al. Rev. Fac. Agron. (LUZ). 2024 41(4): e244141
3-7 |
(5 mm diameter) of active mycelium (three replicates for each strain)
(Prasad et al., 2023). The cultures were kept in incubation (Thermo
Scientic®150) at 26 °C for 10 days. A culture with Pikovskaya
medium without fungus was used as a control. After 10 days, the
content of the asks of each strain was ltered (Whatman® 42). The
extract obtained from each of the strains was measured for soluble
phosphorus content and pH.
Measurement of soluble phosphorus in the extracts of
Trichoderma strains
Was evaluated by the ascorbic acid method (Clesceri et al., 1992).
The absorbances were measured in a spectrophotometer (JENWAY®
6305) at 880 nm phosphorus solubilization was calculated against a
standard curve prepared with KH
2
PO
4
(concentration 1-40 mg.L
−1
).
The data obtained were expressed in mg.mL
-1
.
Study of the ability of Trichoderma strains to solubilize
phosphate in the greenhouse
Inoculum production
Trichoderma strains were transferred to PDA and then incubated
at 26 °C for 7 days to promote the production of conidia. The culture
was immersed in solution of Inex A 0.1% (Cosmocel®) to prepare
concentration to 1.10
-8
CFU.mL
-1
by counting in a Neubauer chamber.
Inoculation of Trichoderma isolates on coee plants of three
varieties
Six months old coee plants of three varieties of C. arabica
(Anacafe, Costa Rica and Marsellesa) were used. The plants were
transplanted into 5 kg pots with fresh soil (table 1) from the coee
plantations, and sterile sand was added in a ratio of 2:1 v/v.
Table 1. Chemical characteristics of the soil used.
Soil pH
Phosphorus
mg.kg
-1
Nitrogen
(%)
Carbon
(%)
Anacafe 4.82 0.73 0.45 3.80
Costa Rica 4.92 0.97 0.42 3.70
Marsellesa 4.98 0.69 0.47 3.80
The design was a randomized block design with 11 treatments [10
Trichoderma strains and 1 control (no fungus)] with 5 replications.
The inoculum of the strains of Trichoderma are added directly
to the substrate and plant roots at a concentration of 1x10
-8
CFU.
mL
-1
according to Souchie et al. (2010). The plants were kept in
the greenhouse for six months, at the end of this time the soluble
phosphorus content of the substrate and the height of the plants were
measured.
Measurement of soluble phosphorus in the substrate of coee
plants
To determine the amount of soluble phosphorus using the
technique of Bray and Kurtz (1945). Phosphorus concentration was
measured with a spectrophotometer (JENWAY ®6305) at 882 nm
and was calculated against a standard curve prepared with KH
2
PO
4
(concentration 0-20 ppm). The obtained data were expressed in
mg.kg
-1
. To calculate the amount of soluble phosphorous use the
following formula: P (mg.Kg): ppm in CC x DM x DV. Where: ppm
in DC: parts per million in the calibration curve, DM: mass dilution
(volume of extractant solution)/(grams of sample) and DV: volume
dilution (capacity)/(sample aliquot added).
Statistical analysis
The trial used a completely randomized design, and data were
analyzed using ANOVA (one-way). Fisher’s test was treated to
separate means. The level of signicance for analysis was p≤0.05. To
know the relationship between the content of solubilized phosphorus
and the pH in the extracts of the Trichoderma strains as well as the
soluble phosphorus content of the substrate with the height of the
plants of the three coee varieties a Pearson evaluation analysis was
performed. All analysis were done using Statistica 8.0 statistical
software.
Results and discussion
Identication of the Trichoderma isolates
Species were assigned by comparing the sequences obtained
with Gen Bank databases. The species identied with 100 % identity
corresponded to T. asperellum (Th40, Th49) and T. harzianum (Th48,
Th53 and Th55); the rest were categorized as related because they did
not have 100 % identity (table 2). Table 2 presents the data obtained
in the analysis del BLASTn algorithm of the National Center for
Biotechnology Information GenBank (NCBI, 2024).
Table 2. Species assignment of Trichoderma strains using BLASTn
algorithm of the National Center for Biotechnology
Information GenBank.
Key Species
Gene bank
accession
numbers
Max
Score
Query
cover
(%)
Identity
Score
(%)
Th12 T. a crassum PQ516492 436 85
90.62
Th24
T. a silva-virgineae PQ514865 726 79
98.78
Th26
T. a virens PQ514858 1074 99
99.34
Th40
T. asperellum PQ514866 1038 100
100
Th48
T. harzianum PQ514859 814 100
100
Th49
T. asperellum PQ514860 1062 98
100
Th53
T. harzianum PQ514861 1109 99
100
Th55
T. harzianum PQ514862 1110 98
100
Th57
T. a asperellum PQ514863 1090 98
99.83
Th78
T. a piramidales PQ514864 1098 99
98.86
The ten evaluated strains had morphological and microscopic
features characteristic of Trichoderma spp. The mycelial color of
all strains starts o white with a cottony appearance and turns light
to dark green after sporulation. Most of them form colonies with
concentric rings. After 5 days of cultivation in PDA medium, the
colonies rapidly grew and matured. Microscopic observation revealed
slightly ovoid conidia arranged in rosettes, apical phialides arranged
in three to four transverse arcs.
Study quantitative of the ability of Trichoderma strains to
solubilize calcium phosphate and aluminum phosphate
.
The soluble phosphorus concentrations of ten Trichoderma
strains evaluated in the culture medium with calcium phosphate
(Ca
2
PO
4
) after 10 days of incubation varied signicantly (p>0.0005),
from 54.42 to 75.73 mg.mL
-1
. In the control treatment, the soluble
phosphorus value was 7.42 mg.mL
-1
. The highest value was detected
in the culture of the strains Th53 (75.73 mg.mL
-1
) and Th48 (74.62
mg.mL
-1
), and the lowest in the strain Th55 (54.42 mg.mL
-1
). The
solubilized phosphorus content was negatively correlated with the pH
(R= -0.78; p< 0.05) (gure 1a). The pH values were 6.35 in the control
and 4.81-3.73 with the dierent Trichoderma treatments (gure 1a).
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Rev. Fac. Agron. (LUZ). 2024, 41(4): e244141 October-December. ISSN 2477-9407.
4-7 |
The soluble phosphorus concentrations of ten Trichoderma strains
in the culture medium with aluminum phosphate (AlPO
4
) after 10
days of incubation varied signicantly (p>0.005), from 7.04 to 22.99
mg.mL
-1
. In the control treatment, the soluble phosphorus value was
7.04 mg.mL
-1
. The highest value was observed in the strains Th57
(22.99 mg.mL
-1
), Th49 (22.90 mg.mL
-1
), and Th40(21.55 mg.mL
-1
).
The strains
Th55 and Th48 presented intermediate values, with no
signicant dierence between them. The lowest solubilization values
were found in Th12 (9.1584 mg.mL
-1
), Th26 (8.52 mg.mL
-1
), and
Th78 (7.04 mg.mL
-1
) (gure 1b). The solubilized phosphorus content
was negatively correlated with the pH (R= -0.47; p< 0.05). The pH
values were 5.21 in the control and 3.38-2.75 with the dierent
Trichoderma treatments. Based on the calcium phosphate (Ca
2
PO
4
)
results, Th53, Th40 y Th26 were selected for the greenhouse study
since they showed the highest solubilization of phosphorus.
A
B
Figure 1. Solubilized phosphorus concentration (bars) and pH
(line) of the extracts of Trichoderma strains (Th12:
T. a crassum, Th24: T. a silva-virgineae, Th26: T.
a virens, Th40: T. asperellum, Th48: T. harzianum,
Th49: T. asperellum, Th53: T. harzianum, Th55:
T. harzianum, Th57: T. a asperellum, Th78:
T. a piramidales) and control in liquid culture
medium with calcium phosphate (a) and aluminum
phosphate (b) after 10 days of incubation. Bars
values represent the mean of three repetitions ±
standard deviation, shared letters in a column do
not indicate signicant dierences when compared
with Fisher's means tests (p<0.05). The line values
represent the mean of three repetitions.
For the Trichoderma
strains analyzed in this study, the values of
phosphorus resulting from the solubilization of aluminum phosphate
were lower than those for calcium phosphate. Zhang et al. (2018) found
the same with strains of Talaromyces aurantiacus and Aspergillus
neoniger. Hernández-Leal et al. (2011) indicated that this can be
explained through the Kps values (solubility product constant or
equilibrium constant of ions in solution) for the dierent phosphates,
the lower the value, the less soluble the compound. Calcium
phosphate has a constant of 2.21x10
-4
, while aluminum phosphate has
a constant of 9.84x10
-21
, making it more dicult to solubilize. The
concentration of solubilized calcium phosphate (Ca
2
PO
4
) in this study
occurred higher than those described for other Trichoderma strains
by Kribel et al. (2019) (up to 12.42 µm.mL
-1
), Tandon et al. (2020)
(up to 70.8 µm.mL
-1
), and Prasad et al. (2023) (48.0-56.0 µm.mL
-1
);
therefore, the strains evaluated in this study have great potential
for use as biofertilizers. Strains T. harzianum (Th48 and Th53)
provided the highest solubilization for calcium phosphate (Ca
2
PO
4
)
and for aluminum phosphate (AlPO
4
); the highest solubilization was
achieved by T. asperellum (Th57), (Th49) and (Th40). With both
sources of insoluble phosphate, a negative relationship with pH was
detected, i.e., the greater the phosphate solubilization, the lower
the pH. Several authors have associated phosphate solubilization in
Trichoderma with the decrease in pH mediated by the production
of organic acids (Bononi et al., 2020); however, the production of
phosphatase enzymes has also been shown. Li et al. (2015) reported
redox, chelation (siderophore formation), and hydrolysis (phytase)-
mediated phosphate solubilization by Trichoderma. These acids are
products of microbial metabolism (lactic acid, diuse acid, ascorbic
acid, isocitric acid, malic acid, citric acid), mainly through oxidative
respiration or by fermentation of carbonaceous substrates (e.g.,
glucose) (Menezes-Blackburn et al., 2016). Although the secretion
capacity and quality of organic acid is basically determined by genetic
regulation, it can also be aected by the excretion of protons in the
assimilation of NH
4
+
(Osorno and Osorio, 2014).
Study of the ability of Trichoderma strains to solubilize
phosphate in the greenhouse
Phosphorus soluble
In general, compared to the control, a signicant increase was
observed in the available phosphorus content in the substrate of
the plants of all varieties inoculated with the Trichoderma strains.
However, the availability of phosphorus and the response of the
plants inoculated with the Trichoderma strains diered among
the three coee varieties. In the plants of the Anacafe variety, the
highest available phosphorus contents were 2.15 mg.kg
-1
and 2.07
mg.kg
-1
, with Th53 and Th48 strains, respectively. These values were
signicantly higher than those in plants inoculated with Th26. The
response pattern of the available phosphorus content of the Costa
Rica and Marsellesa variety plants to the inoculation of Trichoderma
strains was similar; however, the values of soluble phosphorus were
dierent. In both varieties, the highest available phosphorus content
was detected in the plants inoculated with Th48 strain, followed by
the plants inoculated with Th26 and Th53. The available phosphorus
values in the Costa Rica variety were higher (1.80-3.49 mg.kg
-1
)
than those in the Marsellesa variety (1.42-2.97 mg.kg
-1
) and Anacafe
variety (1.13-2.15 mg.kg
-1
) (gure 2a).
Height of coee plants
The heights of the plants inoculated with Trichoderma strains were
dierent in the three coee varieties. In Anacafe plants, the height
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Arias et al. Rev. Fac. Agron. (LUZ). 2024 41(4): e244141
5-7 |
of the plants inoculated with Trichoderma strains was signicantly
greater than the height of the plants without Trichoderma (control).
The height was signicantly higher in those plants inoculated with
Th53 than those inoculated with Th48 and Th26. The heights of
the Costa Rica plants were similar under all treatments (p>0.05). In
the plants of the Marsellesa variety, the heights were signicantly
dierent among all treatments. The greatest height was found in those
inoculated with Th53, followed by Th48 and Th26 (gure 2b).
In the Pearson correlation analyzes a signicant relationship of
phosphorus content with height was only detected in the plants of the
Anacafé variety (R= 0.75; p< 0.05).
A
B
Figure 2. a) Soluble phosphorus in the substrate and b) height
of coee plants of coee of the varieties Anacafe, Costa
Rica and Marsellesa after 6 months of inoculation with
three strains of Trichoderma (Th26: T. a virens, Th48:
T. harzianum, and Th53: T. harzianum) and control
(C). The values are the mean of ve repetitions ± standard
deviation, shared letters in a column indicate no signicant
dierences when compared with the Fisher mean tests
(p<0.05).
Various studies demonstrate that species belonging to the
genus Trichoderma improve the growth response of various crops
(Galeano et al., 2024; Kaissoumi et al., 2024) because Trichoderma
strains can produce phytohormones such as auxins and auxin-like
secondary metabolites, as well as gibberellic acid and ethylene. The
high diversity of Trichoderma species in the rhizosphere of coee
plantations has been demonstrated (Arias and Heredia, 2014; Mulatu
et al., 2022), few studies have addressed the use of strains of this
genus as phosphorus solubilizes. Most of the studies on phosphate
solubilization by Trichoderma species have been performed in
vitro (Tandon et al., 2020; Prasad et al., 2023). In vivo studies with
Trichoderma have been reported with some cultures such as cowpea
(Vigna unguiculata) (Chagas et al., 2016); tomato (Bader et al., 2020);
soybean plant (Galeano et al., 2024). Little research has addressed
the stimulating eect of using phosphate-solubilizing Trichoderma
species on coee plants. In this work, three varieties of coee
plants (Costa Rica, Anacafe, and Marsellesa) were used; though the
Trichoderma strains showed positive responses in the absorption of
phosphorus, the responses of the three varieties of coee plants to the
inoculation of the Trichoderma strains were dierent. The delayed
response is probably due to the coee varieties having a dierent
composition of the exudates that are released, which determines
the interactions that may occur there (Huerta and Holguín, 2019).
In general, the chemical compounds released by plant roots can
interact directly with microorganisms and be directly responsible for
the type of interaction and eect caused. However, the interactions
are extremely complex and a matrix of variables that regulate the
interaction can be generated, to such an extent that there is an overlap
in the chemical language used (Oliveros-Bastidas et al., 2009). The
rhizosphere is an environment rich in nutrients (sugars, organic acids,
amino acids, etc.), probably dierent carbon sources associated with
root exudates could have a signicant eect on the solubilization of
phosphorus that occurs in the rhizosphere (Liebersbach et al., 2004).
It is very important that the selection of bioinoculants involves
native species, since these will likely show better acquisition
of nutrients, better development, and greater modulation in the
protection against pathogens. In the plants of the Anacafe, Costa Rica,
and Marsellesa varieties, the increase in the available phosphorus was
up to 1.42 mg.kg
-1
, 2.52 mg.kg
-1
, and 2.28 mg.kg
-1
, respectively. In the
case of the controls, the increase was limited and was even exhausted
in the plants of the Costa Rica variety. Perea-Rojas et al. (2019) under
controlled conditions using coee plants (C. arabica var. garnica)
inoculated with a consortium of arbuscular mycorrhizal fungi, two
strains of P-solubilizing fungi (Aspergillus niger
and Penicillium
brevicompactum) resulting in an increase of 1.41-3.8 mg.kg
-1
. In
another eld study in three coee plantations, Arias et al., (2023) in
plants (Costa Rica variety) inoculated with a strain of the phosphorus-
solubilizing fungus P. brevicompactum showed a signicant increase
in soil soluble phosphorus, and the benets of this increase were
highlighted in coee bean production.
It is important to emphasize that, even within the same
Trichoderma species, not all isolates can promote plant growth. In
this study, the benecial eect was reected in a higher content of
soluble phosphorus in the substrate, however under these conditions
it was not related to the height of the plants of most coee varieties.
More time will probably be required for this to be reected in this
variable; long-term tests are needed, including studies to analyze
the eect of these fungi on plant survival in the eld. The data show
large dierences in phosphorus content between plants inoculated
and those not inoculated with Trichoderma, which leads us to assume
that under optimal conditions of this nutrient the plant will have a
better development that will subsequently translate into greater crop
productivity and protection against pathogens. In the rhizosphere,
extensive communication occurs between plants and their associated
microorganisms through the exchange and perception of signals. In
this sense, signaling molecules can have a positive or negative eect
on the interaction. However, the composition of organic matter and
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Rev. Fac. Agron. (LUZ). 2024, 41(4): e244141 October-December. ISSN 2477-9407.
6-7 |
the associated biotic and abiotic environment can aect the activities
of Trichoderma (Cano, 2015). It is dicult to predict the outcome
of interactions between plants and benecial soil microorganisms,
since the response of plants to inoculation varies depending on
the functional and biochemical compatibilities of the interaction.
Therefore, further research is required in Mexico to corroborate the
eectiveness of strains with phosphorus solubilizing potential to
develop technological packages for successful application in coee
cultivation. Trichoderma strains may be one of the most important
alternatives to chemical fertilizers that can have negative eects
on human health and the environment. It is recommended that
future studies explore the possibility of promoting these isolates as
biofertilizers to improve phosphorus nutrition in multiple crops.
Conclusions
Two strains of T. harzianum (Th53 y Th48) from coee
plantations are potentially important for plant development, as well
as for the solubilization of insoluble phosphorus. These strains
favored the availability of phosphorus in vitro in the presence of
calcium phosphate (Ca
2
PO
4
) as well as a higher phosphorus content
in the substrate of the three coee varieties (Anacafe, Costa Rica and
Marsellesa). For its part, T. harzianum (Th48) promoted the height
of the Anacafé and Marsellesa varieties. The T. asperellum strains
(Th40, Th49, and Th57) were the best at solubilizing aluminum
phosphate (AlPO
4
); this was only evaluated under in vitro conditions,
so it is recommended to continue with the tests in coee seedlings.
It would be interesting to evaluate other coee variables, as well as
more phosphorus-solubilizing strains and evaluate other agronomic
variables. It is also recommended to evaluate plant phosphorus
acquisition through whole-plant phosphorus analysis. Field survival
tests are also necessary to evaluate the activity of these fungi in
dierent environmental conditions. It also deserves further research
to evaluate the eect of the application of Trichoderma consortia and
with consortia of other microorganisms such as mycorrhizae on the
mechanisms of phosphor mobilization. The results presented here are
a basis for the development of future study aimed at the use of native
and properly identied Trichoderma strains in coee farming and
inuencing the use of biofertilizers as an environmentally friendly
strategy that also favors the production of organic coee since it has
a better position in the market and thus favors the economy of coee
producers in the region. It is suggested to continue the exploration of
the selection of strains with high potential to solubilize both calcium,
aluminum and iron phosphate, continue testing on application
methods and promote integration with other sustainable practices.
Funding
This study was funded by COVEICYDET Project 131627.
Acknowledgments
The authors thank Alondra Guadalupe Martínez Santos and María
del Rosario Gregorio Cipriano for their support in the molecular
identication of the strains.
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