ORIGINAL RESEARCH

published: 26 July 2022

doi: 10.3389/fpls.2022.897668

Biological Control and Plant Growth

Promotion Properties of Volatile
Organic Compound-Producing
Antagonistic Trichoderma spp.
Jin Ho Joo 1† and Khalid Abdallah Hussein 1,2*†
1
Soil Biochemistry Lab, Department of Biological Environment, Kangwon National University, Chuncheon, South Korea,
2
Botany and Microbiology Department, Faculty of Science, Assiut University, Asyut, Egypt

Trichoderma is environmentally vital due to their plant growth-promoting effects (such

as enhancement of nutrients supply, suppression of plant pathogens, and promotion
of plant defense). Biogenic volatile organic compounds (VOCs) are diverse chemical
substances emitted by Trichoderma spp. The potential role of VOCs in biological control
and plant growth promotion has recently been recognized. Here, the Trichoderma-VOCs’
performance for plant growth promotion and suppression of plant pathogens are
evaluated. We further investigated VOC emission profiles of T. harzianum using
Edited by: GC–MS. The Trichoderma-VOCs exhibited significant (p < 0.05) antifungal properties
Ying Ma,
University of Coimbra, Portugal against all tested pathogenic fungi. T. atroviride-VOCs showed a decisive inhibition
Reviewed by: of Alternaria panax, Botrytis cinerea, Cylindrocarpon destructans, and Sclerotinia
Rosa E. Cardoza, nivalis. The germinating seeds demonstrated growth enhancement in the presence
Universidad de León, Spain
Milena Cotoras,
of Trichoderma-VOCs emitted by different strains. Low levels of cyclopentasiloxane,
University of Santiago, Chile decamethyl, cyclotetrasiloxane, and octamethyl were found in T. harzianum KNU1 strain
*Correspondence: whereas cyclopentasiloxane, decamethyl, cyclotetrasiloxane, and octamethyl showed
Khalid Abdallah Hussein higher emission levels as Si-containing compounds. The results reveal the potentiality
[email protected]
of VOCs as a biocontrol resource against deleterious rhizosphere microorganisms and
† These authors have contributed underline the importance of Trichoderma-VOCs emissions in regulating plant growth
equally to this work
and development.
Specialty section: Keywords: volatile organic compounds, plant growth-promotion, fungi, environment, agriculture
This article was submitted to
Plant Symbiotic Interactions,
a section of the journal
Frontiers in Plant Science
INTRODUCTION
Received: 19 March 2022 The reduction of fertilizers and fungicides used in the agricultural field is an insistent demand to
Accepted: 20 June 2022 maintain the ecosystem and improve sustainable agriculture. Currently, beneficial microorganisms
Published: 26 July 2022
are progressively being used as inoculants for biofertilization and biocontrol. Seed or seedling
Citation: treatment with both biofertilizers and biocontrol systems has been used to improve the growth
Joo JH and Hussein KA (2022)
of plant crops and minimize the effects on the environment (Kumar et al., 2022). Trichoderma
Biological Control and Plant Growth
Promotion Properties of Volatile
is a filamentous fungus that is one of the most widely studied genera with many applications
Organic Compound-Producing in agriculture and the environment (Mukherjee et al., 2013; Hussein et al., 2020). Several
Antagonistic Trichoderma spp.. Trichoderma species possess the ability to suppress plant diseases along with improving plant
Front. Plant Sci. 13:897668. growth and productivity via overlapping modes of action such as enhanced nutrient efficiency
doi: 10.3389/fpls.2022.897668 (Zin and Badaluddin, 2020), induced systemic resistance (Farag et al., 2013; Tahir et al., 2017),

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

and mycoparasitism (Mukherjee et al., 2022). In agriculture, the plant-pathogens relationship. Little is known about plant
Trichoderma species are potent biocontrol agents and are growth promotion and resistance offered by VOCs-producing
often applied to soils to control soil-borne pathogens and PGPF (Tilocca et al., 2020). In this context, this study aimed to
increase crop yields worldwide. More than 250 Trichoderma- detect the plant growth promotion and the biological control
based formulations were estimated in India (Mendoza-Mendoza properties of the airborne chemicals released from Trichoderma
et al., 2015). Moreover, Trichoderma species possess a natural rhizosphere strains.
resistance to many chemicals and fungicides used in agriculture,
therefore, they are readily integrated into agricultural practices MATERIALS AND METHODS
(Liu and Zhang, 2015; Ons et al., 2020). Trichoderma species
are fertile producers of many metabolites with antibacterial, Sampling and Fungal Cultures
antifungal and anticancer properties (Khan et al., 2020). Trichoderma strains were isolated from the Korean ginseng
Trichoderma spp. release a wide spectrum of volatile organic (Panax ginseng) and pine (Pinus koraiensis) soils (rhizosphere)
compounds (VOCs). It has been revealed that Trichoderma from different localities within Kangwon-do province,
VOCs play a vital role in inter-species communication and Chuncheon, Korea. The soil samples were precisely taken
can protect plants against some soil-borne pathogens. Although in zip bags and kept at 5◦ C. The Trichoderma strains were
the signaling network between microbes and plants has isolated and purified on potato dextrose Rose-Bengal agar
been extensively studied in the recent two decades, the role (PDA amended with 50 µg/ml Rose Bengal) and grown at 27◦ C
of rhizomicrobial VOCs in regulating and enhancing plant (Heydari and Pessarakli, 2010). Among forty-five isolates, only
growth needs more investigations. Yamagiwa et al. (2011) first 10 strains were selected based on their antagonism potentiality
reported about the role of VOCs produced by plant growth- as bioagents through dual culture technique. Trichoderma spp.
promoting fungi (PGPF) in growth promotion. They introduced were identified according to the cultures’ features and the direct
Talaromyces wortmannii as a new PGPF, having growth microscopic examination (Domsch et al., 1980; Moubasher,
promotion traits on several plants, such as Arabidopsis thaliana, 1993). The universal primers ITS1: 5′ TCC GTA GGT GAA
Brassica campestris, and Phaseolus vulgaries. Microbial volatiles CCT GCG G 3′ and ITS4: 5′ -TCCTCCGCTTATTGATATGC-
play a proper role in the agricultural environment by promoting 3′ were used for fungal amplification (Herlemann et al., 2011).
plant growth and inducing the resistance mechanism against The PCR amplifications were performed in a thermal cycler
plant pathogens, without any harmful effect on the environment for 30 cycles at 94◦ C for 1 min for DNA denaturation, primer
(Schulz-Bohm et al., 2017; Tilocca et al., 2020). Recently, the annealing for 30 s at 56◦ C, and primer extension for 1 min
role of volatile materials from rhizobacteria in regulating plant at 72◦ C. The PCR products were sequenced at Macrogen
growth and development has been widely considered. Ryu et al. Inc. The partial gene sequences were matched with the full
(2003) noticed that a blend of VOCs released from specific sequences presented in the GenBank database using the BLAST
plant growth-promoting rhizobacteria (PGPR) strains promoted search (NCBI). The phylogenetic tree structure was shaped
the growth of the seedlings of A. thaliana. Gutiérrez-Luna using the online tool www.phylogeny.fr (PhyML), and the
et al. (2010) also mentioned that airborne chemicals from tree visualization was done using TreeDyn. The investigated
some Bacillius sp. strains have a growth enhancement effect. identified Trichoderma strains have been deposited in the Korean
Farag et al. (2013) found that bacterial volatile compounds are agricultural culture collection (KACC) as shown in Table 1. The
involved in plant growth promotion during their assessment in fungal phytopathogenic isolates Alternaria panax, Botrytis
vitro for rhizobacteria. Trichoderma spp. are known as PGPF cinerea, Cylindrocarpon destructans, Fusarium oxysporum,
having the ability to promote plant health and to compete Sclerotinia nivalis, and S. sclerotiorum were obtained from
against pathogenic microbes (Bissett et al., 2015; El-Maraghy KACC, Jeonju, Korea. The pathogenic fungi were maintained on
et al., 2020a). The genus Trichoderma comprises numerous 2% malt extract agar (MEA) grown at 24◦ C and stored at 4◦ C
species (Bitas et al., 2013) which are ubiquitously existent in before further investigation.
agricultural soils and forests, where they intensively interact with
plant roots and rhizosphere microorganisms (Contreras-Cornejo Siderophores Biosynthesis Assay
et al., 2016). A profound understanding of the Trichoderma The CAS (Chrome azurol S) liquid assay was conducted
properties, including metabolic activity and the interaction according to Schwyn and Neilands (1987) to detect the
with plants and other microbes, can guarantee its effective use biosynthesis of the siderophores quantitatively. The solution
in agriculture (Tyskiewicz et al., 2022). The interest in the was amended to pH 6.8 with a 0.1 M Pipes buffer (Prod. No.
application of Trichoderma is growing fast due to their direct P1851, Sigma). The fungal cultures were inoculated into liquid
and indirect biocontrol activity against a wide range of soil-borne deferrated Czapek’s medium and incubated at 25◦ C in a rotary
disease (Tyskiewicz et al., 2022). Several studies investigated shaker (ICB-S0420, Co., Ltd. China) at 150 rpm till they reached
Trichoderma’s potential to control soil phytopathogens through the maximum stationary phase (5 × 107 spores/ml). Then 0.5 ml
various complicated mechanisms, such as mycoparasitism, culture supernatant was added to 0.5 ml CAS solution and 10 µl
competition for nutrients and space, the degradation of pathogen 5-sulfosalicylic acid 0.2 M (as shuttle solution) and was mixed
cell walls, and induction of plant resistance, VOCs also seem carefully. The mixtures were settled for 10 min. The color change
a promising approach. Moreover, most studies have been was determined by absorbance (A630 ) for the vanish of blue color
interested in VOCs released from PGPR and their impact on using a UV-VIS spectrophotometer (Agilent Inc., United States).

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

TABLE 1 | Trichoderma strains isolated from rhizosphere soils and their identification.

Strains Max identity (%) Strain of closest identity Identification Accession No.

T-KNU1 98% Trichoderma harzianum BGP Trichoderma harzianum KNU1 KACC 42249
T-KNU4 98% Trichoderma reesei SU_Solok Trichoderma reesei KNU4 KACC 40557
T-KNU10 98% Trichoderma harzianum BCS8A Trichoderma harzianum KNU10 KACC 40558
T-H22 100% Trichoderma harzianum BGP115 Trichoderma harzianum H22 KACC 40779
T-24 100% Trichoderma atroviride IMI 206040 Trichoderma atroviride 24 KACC 40776
T-27 100% Trichoderma koningii T-400 Trichoderma koningii 27 KACC 40779
T-19 100% Trichoderma virens Gv29-8 Trichoderma virens 19 KACC 40780
T-28 100% Trichoderma longibrachiatum H9 Trichoderma longibrachiatum 28 KACC 40798
T-P22 99% Trichoderma Pleuroticola GJS Trichoderma Pleuroticola P22 KACC 42583
T-18 99% Trichoderma asperellum IIPR-Ta10 Trichoderma asperellum 18 KACC 43821

An un-inoculated culture medium was served as blank and Indole-3-Acetic Acid Assay
the un-inoculated culture medium containing CAS and shuttle A total of ten different Trichoderma isolates were quantitatively
solution was served as a reference. The siderophore production tested to biosynthesis IAA according to Brick et al. (1991).
was evaluated based on the following Equation (1): Trichoderma spp. were grown in Czapek-Dox broth amended
with tryptophan (1000 µg ml−1 ), as a nitrogen source, instead
(Ar − As)/Ar 100 = % siderophores units of NaNO3 . In a conical tube (50 ml), 10 ml of the liquid bioassay
media was inoculated by actively growing hyphae, incubated at
27◦ C, and agitated at 150 rpm. After 7 days, each Trichoderma
where As is the absorbance rate of the sample, and Ar is the culture was centrifuged at 16,099 × g for 15 min. Salkowski’s
absorbance rate of the reference. The experiment was achieved reagent was prepared (10 mM FeCl3 in 35% HClO4 ) to detect
in triplicates and average values were used. indole derivatives. About 1 ml of the supernatant was added
to the same volume of the reagent and kept for 30 min in
Phosphate Solubilization Assay dark conditions. The optical density was detected using UV
The precipitated Ca3 (PO4 )2 on Pikovskaya’s agar media (g L–1 ; spectroscopy (Hitachi U-2900) at 530 nm. A standard curve of
(NH4 )2 SO4 , 0.5 g; Ca3 (PO4 )2 , 5 g; glucose, 10 g; NaCl, 0.2 g; KCl, IAA concentrations was designed to evaluate the corresponding
0.2 g; MgSO4 .7H2 O, 0.1 g; FeSO4 , 0.002 g; MnSO4 , 0.002 g; yeast concentration of IAA released by each Trichoderma strain in the
extract, 0.5 g; agar, 15 g) was used for the qualitative detection of bioassay media.
the phosphate-solubilizing Trichoderma (Pikovskaya, 1948). All
Trichoderma spp. were grown at 27 ◦ C as surface-culture in 50 ml Pathogen-Fungicide Assay
conical tubes containing 20 ml of Czapek-Dox broth fortified The inhibitory effects of fenhexamid (50%, Indofil) and
with 0.1% Tween 80. After 1 week, the cultures were vortexed mancozeb (75%, Indofil) on the linear growth of pathogenic
for 1 min and the inocula size was finally adjusted to reach an fungi were investigated using the dilution-plate method. The
optical density of 0.3 McFarland Standard (corresponds to ∼5 × fungicide solution was prepared in sterile H2 O and added to
105 Cells/ml) using the spectroscopy (UV-VIS, Hitachi U-2900) the autoclaved PDA (∼50◦ C) to reach the final concentration
at 600 nm (McFarland, 1907). The solidified Pikovskaya’s agar of 20 µg ml−1 for each chemical fungicide (Abdoon et al.,
media was inoculated by Trichoderma spp. using a bacteriological 2011). The mixture was decanted into Petri dishes (90 × 15 mm)
isolation loop. The cultures were incubated for 7 days at 27◦ C before solidification. Mycelial agar disc (5 mm) was cutoff from
and the developed clear zone was detected (Pingale and Virkar, the rim of the freshly growing culture of pathogenic fungi
2013). Phosphate solubilization was detected quantitatively using and inoculated on the fungicide-amended media surface. The
Pikovskaya’s broth (pH was adjusted to 7.0). About 10 ml of pathogen-fungicide assays were achieved in three replications;
Pikovskaya’s solution was inoculated using an isolation loop all the cultures were incubated at 24◦ C for 10 days in darkness.
full of Trichoderma spore suspension to unify the inoculi size; The colonies’ diameters were measured and the inhibition indices
cultures were kept at 27◦ C for 7 days (Pikovskaya, 1948). were evaluated in comparison to control (fungicide-free cultures)
Then the cultures were centrifuged at 16,099 × g for 10 min. (Messgo-Moumene et al., 2015).
The supernatant was filtrated using Whatman No. 2 filter to
remove the color impurities. In 24 well plates, equal volumes of Pathogen-Trichoderma VOC-Exposure
Barton’s reagent and supernatant were added and left for 10 min, Bioassay
The color intensity was assessed using a colorimeter system The Trichoderma isolates were inoculated in malt extract agar
(Nanodrop Biotech, United States) at wavelength 430 nm, and the media (MEA) and incubated at 27◦ C. One day later, the
soluble phosphorus quantity was detected using a standard curve covers of the Petri dish were exchanged for the bottoms of
(Pingale and Virkar, 2013). the 3-day-old PDA cultures of the phytopathogenic fungi. The

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

FIGURE 1 | Colony appearance of the different selected Trichoderma spp. isolates on PDA (7 days). (A) Trichoderma harzianum KNU1; (B) T. reesei KNU4; (C) T.
harzianum KNU10; (D) T. harzianum H22; (E) T. atroviride 24; (F) T. koningii 27; (G) T. virens 19; (H) T. longibrachiatum 28; (I) T. Pleuroticola P22; (J) T. asperellum 18.

TABLE 2 | Plant growth-promoting activities of the selected rhizosphere Trichoderma sp. strains.

Strains Siderophore IAA production Phosphate solubilization

units (%) µg ml−1 µg ml−1

Trichoderma harzianum KNU1 29.54 ± 1.95bc 21.25 ± 1.72bc 71.1 ± 5.11b

Trichoderma reesei KNU4 31.73 ± 4.31c 53.63 ± 15.03a 45.02 ± 4.01bc
Trichoderma harzianum KNU10 33.48 ± 3.18c 59.10 ± 6.12a 21.52 ± 2.19c
Trichoderma harzianum H22 40.26 ± 5.11b 21.48 ± 15.39bc 12.625 ± 1.89c
Trichoderma atroviride 24 43.11 ± 3.83b 18.97 ± 13.63bc 12.16 ± 1.14cd
Trichoderma koningii 27 91.68 ± 5.33a 24.60 ± 13.10bc 106.17 ± 6.11a
Trichoderma virens 19 30.2 ± 2.98c 36.60 ± 17.04ab 152.155 ± 6.89a
c bc
Trichoderma longibrachiatum 28 33.48 ± 2.87 24.51 ± 10.23 22.485 ± 2.33c
d bc
Trichoderma Pleuroticola P22 20.69 ± 2.21 19.69 ± 3.67 19.04 ± 2.15c
b c
Trichoderma asperellum 18 49.67 ± 2.53 10.94 ± 9.35 2.06 ± 1.91d

Same letters within a column are not significantly different at P < 0.05.

two halves/cultures of the plate were wrapped together using Plant-Trichoderma VOC-Exposure
impermeable parafilm tape and incubated for 10 days at 24◦ C. Bioassay
The linear growth of the six pathogenic fungi was detected. The Seeds of Raphanus sativus L. (radish plant) were surface sterilized
bioassay was achieved in triplicates. The controls were inoculated by soaking in 70% ethanol for 3 min, rinsed in sterile distilled
only with the fungal pathogens. All the bioassay procedures water (three times), and placed on Petri dishes (I-plates; Atekuto)
were conducted under light-limited conditions to normalize the containing Murashige and Skoog (MS) salt solid medium (pH
Trichoderma sporulation (Di Lelio et al., 2021). Observations on was adjusted to 5.7) on one side, and a small Petri dish (35
the antifungal activities of Trichoderma VOCs and the inhibition × 10 mm) containing newly growing Trichoderma on PDA was
percentage were reported according to the method of Messgo- put on the other side. A total of five seeds per plate were
Moumene et al. (2015) as shown in the following Equation (2): transferred to the MS medium side (Murashige and Skoog, 2006).
Uninoculated PDA in a small Petri dish was set for control.
C1 − C2 /C1 ×100 = Inhibition percentage (%)
The I plates were sealed with Parafilm tape and incubated
where, C1 is the colony surface area of the uninhibited pathogenic randomly in the growth chamber (DASOL; Scientific Co., Ltd.,
fungus in the control, while C2 is the colony surface area of the Korea) which was adapted to 25◦ C and 12:12-h light: dark
pathogenic fungus in the dual culture. (LD) conditions.

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

Volatile Metabolites Analysis by T. reesei KNU4, T. harzianum KNU10, T. harzianum H22, T.

Headspace GC–MS atroviride 24, T. koningii 27, T. virens 19, T. longibrachiatum
T. harzianum KNU1 strain was cultured in 10 ml SPME glass 28, T. Pleuroticola P22, and T. asperellum 18) that showed
headspace vials (Supelco, Bellefonte, St Louis, United States) dual antagonistic effects and different morphological features
containing PDA medium and incubated at 27 C for 7 days. were identified and investigated for plant growth-promoting
The vials were sealed with a duratool-crimp cap fitted with a traits, e.g., siderophore production (Figure 1, Table 1). The
silicon/Teflon septum that was previously adopted at 100◦ C for selected rhizosphere Trichoderma strains were varied in terms
30 min. Headspace samples taken from sterile PDA served as a of their plant growth-promoting traits. For example, the T.
negative control. The analysis of Trichoderma VOCs was carried koningii strain showed the maximum siderophore production
out by an Agilent SPME and GC-MS (GEOL and 7890A GC (91.7% siderophore units) and solubilized 106.1 µg ml−1 of
MSD JP/JMS-Q1050GC) equipped with a split-less injection and inorganic phosphate. However, T. asperellum produced 49.7%
a capillary DB-WAX GC column (30 × 0.32 mm and 0.5 µ film siderophore units and solubilized only 2.1 µg ml−1 of inorganic
thickness). The injection port temperature was set at 250◦ C and phosphate. The lowest siderophore production was by T.
the column oven temperature was programmed in the range of pleuroticola P22 strain which produced only 20.7% siderophore
50–250◦ C (5◦ C min−1 ), then elevated to 300◦ C (5◦ C min−1 ), units (Table 2). The evaluation of the IAA biosynthesis by
ending with 5 min isothermal at 300◦ C. Helium (1 ml min−1 ) the selected Trichoderma revealed that T. reesei KNU4 and
was the carrier gas and all mass spectra were detected at 70 eV. T. harzianum KNU10 strains produced high amounts of IAA
The chemical constituents were identified by matching the mass (53.6 and 59.1 µg ml−1 , respectively) whereas all the other
division patterns of the constituents/components to those of the strains produced only <25 µg ml−1 of IAA except T. virens
WILEY reference standards data. 19 which produced 36.6 µg ml−1 . The free living-Trichoderma
strains isolated from the rhizosphere showed a high capability
Statistical Analyses to mobilize insoluble phosphate. T. virens 19 and T. koningii
The experimental design was randomized. Each treatment was strains were potent in phosphate solubilization. They solubilized
carried out with at least three replications and compared with 152.1 µg ml−1 and 106.1 µg ml−1 , respectively. This result was
the negative control. The data were analyzed by the variance followed by T. harzianum KNU1, which also showed strong
using Duncan’s multiple range test (ANOVA) and p-value was phosphate solubilization and liberated 71.1 µg ml−1 of inorganic
evaluated. The bars represented the mean values and standard phosphate (Table 2). Generally, fungi synthesize siderophores
deviation (SD). of hydroxamate-type, such as ferrichromes, coprogens, and
fusarinines. Trichoderma can inhibit the activity and growth
of target soil pathogens by depletion of iron sources in a
RESULTS AND DISCUSSION common niche (Harman et al., 2004). Several representatives of
Trichoderma are characterized by the capability to synthesize
Recently, rhizospheric microorganisms got great attention phytohormones and phytoregulators, including indole-3-acetic
because of their ability to produce strong antimicrobial acid (IAA), which regulates plant development (Ozimek et al.,
volatile compounds. Many rhizosphere fungi are known to 2018; Jaroszuk-’Sciseł et al., 2019; Alfiky and Weisskopf, 2021).
produce volatile organic compounds, which include terpenes,
hydrocarbons, flavonoids, alkaloids, and cyclohexanes. Many of
these compounds displayed anti-microbial activities and fuel Bioactivity of VOCs Produced by
production (Naik, 2018; Hussein et al., 2020). The endophytic Trichoderma
Phoma sp. emitted volatile compounds such as caryophyllene, With the constant contact of plants with various phytopathogens,
some sesquiterpenoids, alcohols, and naphthalene derivatives, and the increased resistance of these pathogens to chemical
which completely suppressed species belonging to Sclerotinia, pesticides, developing biological protection alternatives became
Ceratocystis, and Verticillium (Strobel et al., 2011). The volatile an inevitable demand. Among non-pathogenic microbes,
compounds of the endophytic fungus Colletotrichum truncatum Trichoderma species seem to be the best candidates in the green
showed a strong inhibitory effect on Fusarium sclerotiorum biotechnology due to their biofertilization and biostimulatory
(Kumar and Kaushik, 2013). properties. Most Trichoderma species belong to plant growth-
promoting fungi that can produce phytohormones and the
Trichoderma Plant Growth-Promotion 1-aminocyclopropane-1-carboxylate (ACC) deaminase enzyme
Properties (Tyskiewicz et al., 2022). Trichoderma can be identified as the
Trichoderma is a genus of filamentous fungi, ubiquitous around genus with the highest biocontrol potential due to many isolated
the world, usually colonizing decaying wood and other forms antifungal bioactive compounds and the biostimulation potential
of plant debris (Howell, 2003). Trichoderma is a dominant of Trichoderma (Thambugala et al., 2020; Vishwakarma et al.,
mycobiome component of various soil ecosystems (such as 2020; Rush et al., 2021). In this study, the pathogen Trichoderma
forests, prairie, farmland, salt marshes, and deserts) in all climatic VOC-exposure bioassays exhibited significant suppressions to
areas, including temperate and tropical, and the Antarctica the growth of phytopathogens. The VOCs from T. Pleuroticola
regions (Kamala et al., 2015; Ghorbanpour et al., 2018). In this P22 showed very high inhibition indices against all tested ginseng
study, ten rhizosphere Trichoderma strains (T. harzianum KNU1, root-rot fungi. T. atroviride and T. koningii strains exhibited

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

TABLE 3 | The antifungal effect of Trichoderma spp. and chemical fungicide against the phytopathogenic fungi.

Antifungal agents Alternaria Botrytis Cylindrocarpon Fusarium Sclerotinia S. sclerotiorum

panax cinerea destructans oxysporum nivalis

Trichoderma harzianum KNU1 67.5 ± 3.7a 54.4 ± 2.5a 84 ± 6.6a 63.9 ± 1.96a 57.1 ± 14.14ab 54.7 ± 6.78a
Trichoderma reesei KNU4 25.4 ± 6.2c 40.4 ± 2.5ab 28 ± 28.3bc 59.1 ± 0.98a 34.7 ± 0.83c 29.4 ± 16.64c
Trichoderma harzianum KNU10 40.4 ± 5a 16.7 ± 6c 46 ± 8.5a 39.6 ± 0.98b 26.5 ± 12.48bc 20 ± 1cd
a a a c ab
Trichoderma harzianum H22 86.8 ± 1.2 72 ± 5.7 66.7 ± 2.5 20.1 ± 4.91 47.7 ± 7.49 45.3 ± 2.5ab
a a a b a
Trichoderma atroviride 24 100 ± 1 66.7 ± 7.4 100 ± 1 43.8 ± 8.84 64.1 ± 5.82 55.9 ± 4.16a
a a a b a
Trichoderma koningii 27 85.1 ± 1.2 100 ± 1 45.6 ± 2.5 35.4 ± 2.95 58.8 ± 16.64 42.9 ± 5.82bc
a a a a a
Trichoderma virens 19 90.35 ± 1.2 49.1 ± 12.41 56 ± 5.7 65.3 ± 1.96 59.4 ± 0.83 41.8 ± 0.83bc
a a a a c
Trichoderma longibrachiatum 28 100 ± 2 46.5 ± 1.24 54 ± 19.8 66 ± 0.98 42.4 ± 1.66 77.06 ± 0.83a
a ab a a a
Trichoderma Pleuroticola P22 83.3 ± 1.2 61.4 ± 7.44 70 ± 7.4 82.6 ± 10.8 75.9 ± 0.83 85.3 ± 4.16a
a a a c c
Trichoderma asperellum 18 100 ± 3 54.4 ± 12.41 100 ± 3 22.9 ± 0.98 34.7 ± 0.83 41.8 ± 4.16b
Fenhexamid (20 µg ml−1 ) 30 ± 1d 2 ± 1e 20 ± 2e 13 ± 0.58d 2 ± 1e 13 ± 2d
Mancozeb (20 µg ml−1 ) 45 ± 2c 2 ± 0.58e 35 ± 0.6c 10 ± 1d 2 ± 0.6e 19 ± 0.58d

Same letters are not significantly different at P < 0.05.

FIGURE 2 | Antifungal activity of the VOCs of Trichoderma strains against ginseng-root rot fungi in comparison to fungicides (20 µg ml−1 ) on B. cinerea, A. panax, C.
destructans, F. oxysporum, S. sclerotiorum, and S. nivalis. The antifungal indices were detected after 10 days.

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

FIGURE 3 | The development of Raphanus sativus L. seeds in Murashige and Skoog solid medium in response to plant growth-promoting Trichoderma-volatiles
exposure of (A) Un-inoculated control; (B) T. harzianum KNU1; (C) T. reesei; (D) T. harzianum KNU10; (E) T. harzianum H22; and (F) T. atroviride. The I-plates were
sealed with Parafilm tape and incubated randomly in the growth chamber which was adapted to 25◦ C and 12:12-h (LD) cycle. (G) Fresh weight average induced in
radish seeds by the VOCs released by different Trichoderma strains.

almost similar plant growth-promoting effects; however, the be greater than that of T. koningii-VOCs. T. atroviride-VOCs
inhibition (antifungal index) of T. atroviride-VOCs appeared to and T. asperellum-VOCs only exhibited the highest antifungal

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

FIGURE 4 | Shows the I-plate system used for assessing plant growth promotion in response to plant growth-promoting Trichoderma-volatiles exposure. This setup
allows only volatile compounds to be exchanged, while preventing any diffusion of non-volatile metabolites through the medium.

properties (100% inhibition) against the key phytopathogen B. cinerea. Consequently, Trichoderma-VOCs can exhibit more
of ginseng plant C. destructans. The lowest antifungal effect of benefits in the terms of biological control and plant growth
all Trichoderma spp. VOCs was detected in the T. harzianum promotion over sustainable agriculture.
KNU10 strain which showed only 17% inhibition against B. In addition to the composition of the nutrient media,
cinerea and 20% inhibition against S. sclerotiorum. The volatile the composition of the gaseous emission is another effective
chemicals emitted by T. atroviride exhibited complete inhibition factor for the expedient growth and development of plants
(100%) twice, against A. panax and C. destructans. Similarly, the (Yamagiwa et al., 2011). Different volatile compounds are present
T. asperellum-VOCs caused 100% growth inhibition against the in this gaseous emission, such as nitrogen, carbon dioxide,
same phytopathogens. The airborne chemicals released by T. oxygen, and other types of volatile components produced by
koningii and T. longibrachiatum showed 100% inhibition of B. surrounding organisms, involving the plant itself (Naznin et al.,
cinerea and A. panax, respectively. The highest antifungal activity 2013). In this study, Trichoderma-VOCs significantly enhanced
against A. panax (100% inhibition) was shown by T. atroviride, the early germination of R. sativus seeds. Plant-Trichoderma
T. longibrachiatum, and T. asperellum. The lowest antifungal and pathogens-Trichoderma VOC-exposure bioassays revealed
index was detected in the B. cinerea pathogen (16.7% inhibition) (39.8–210%) increase in plant biomass and (16.7–100%)
by T. harzianum KNU10. The volatile compounds blend emitted suppression of phytopathogens, respectively. The VOCs emitted
by T. harzianum KNU10 showed the lowest antifungal activity. by T. harzianum H22 were associated with the greatest R. sativus
It inhibited the mycelial growth of B. cinerea, S. nivalis, and S. growth promotion. It increased the fresh weight of the early
sclerotiorum by inhibition percentages of only 16.7, 26.5, and germinated seeds by more than 200%. However, the volatile
20%. The chemical fungicide Mancozeb was found to be more compounds emitted by T. reesei, T. virens, and T. longibrachiatum
effective than fenhexamid, showing 2–45% inhibition indices showed relatively low seedling development. They increased the
for all tested phytopathogens. However, fenhexamid showed fresh weight of early germinated seeds by only 39.8%, 19.4%,
inhibition indices between 2 and 30% at the same concentrations and 6.5%. In this investigation, T. virens and T. longibrachiatum
(Table 3). Generally, the most frequently recommended field did not show a significant increase in the fresh weight of R.
concentration of fungicides is between 3 and 35 µg ml−1 sativus early germinated seeds (Figure 3). However, these two
(Abdoon et al., 2011; Aleksova et al., 2021). The two chemical strains showed the highest antifungal index (inhibition indices)
fungicides used in this study exhibited inhibition indices of against the ginseng root-rot fungus A. panax. Although the
<50% in mycelial growth at 20 µg ml−1 . However, the inhibition T. Pleuroticola strain showed a relatively low plant growth-
index of Trichoderma-VOCs on the fungal growth reached 100% promoting effect, it produced high pathogen-inhibiting VOCs.
several times (Figure 2). For example, both T. atroviride-VOCs Similarly, the germinating seeds exposed to VOCs from T.
and T. asperellum-VOCs achieved 100% inhibition indices twice Pleuroticola P22 showed only a 47% increase in seedlings’
against the tested phytopathogens (Table 3). Also, the lowest fresh weight. However, T. Pleuroticola P22 showed very high
inhibition percentage of T. pleuroticola-VOCs was 61.4% against inhibition indices against all tested pathogenic fungi. The VOCs

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

TABLE 4 | GC-MS analysis showing the VOCs profile of PGPF strain Trichoderma harzianum KNU1.

Peak Compound RT Concentration

No. (min) (%)

15 Acids 2-Pentenoic acid, 2-ethyl-, methyl ester, (e)- 11.47 0.03

71 Khusilic acid 23.59 0.15
10 Alcohols 1-Butanol, 2,2-Dimethyle 8.195 0.02
22 2-Amino-1-Phenyl-1-Propanol 16.41 0.08
11 Aldhydes 2,4-Hexadienal 9.45 0.07
29 Benzaldehyde, 2,4-bis(trimethylsiloxy)- 16.95 0.56
8 Alkenes 2,4-Hexadiene, 2,5-dimethyl- 6.82 0.03
12 Heptane, 3-methylene- 10.03 0.03
16 1,3-Cyclohexadiene, 2-methyl-5-(1-methylethyl)- 11.53 0.03
17 1,5-Cyclooctadiene, 1,5-dimethyl- 11.75 0.16
32 2-Methyl-6-methylene-1,7-octadiene 17.17 6.97
36 1H-Cycloprop[e]azulene, 1a,2,3,4,4a,5,6,7b-octahydro-1,1,4,7-tetramethyl 17.45 1.32
38 Bicyclo[7.2.0]Undec-4-Ene, 4,11,11-Trimethyl-8-Methylene 17.54 0.73
43 2,5-Dimethyl-3-vinyl-1,4-hexadiene (Santolina triene) 18.11 1.81
44 Cis-1,3-Pentadiene 18.17 1.40
46 8-Isopropyl-1-Methyl-5-M Ethylene-1,6-Cyclodecadiene 18.45 3.57
5 Amines 2-Hexanamine, 4-methyl- 2.68 0.09
23 Phenylpropropanolamine 16.45 0.11
75 Methylamphetamine 28.77 0.03
24 Esters 1,5-Dideuteriohexane 1-Pentyl-2,2-D2 Acetate 16.51 0.47
62 Hexadecamethyl Octasiloxane 20.73 0.04
74 2-Chlorophenyl 6-(4-Morpholinyl)Hexyl Ether 28.45 0.08
77 Ethyl 2-Methyl-3-Oxo-3-(2-Pyridinyl)Propanoate 29.61 0.01
78 Diethyl 7-Amino-6-Oxo-6h-Benzo [C]Chromene-8,9-Dicarboxylate 30.14 0.04
79 Ethyl 2-Phenyl-4-Phenylthio Methylthiazole-5-Carboxylate 30.17 0.01
4 Hydrocarbons Acetonitrile-d3 2.63 0.04
6 2,3-Pentadiene, 2,4-dimethyl- 4.72 0.06
30 Tricyclo[4.4.0.02,7]dec-3-ene, 1,3-dimethyl-8-(1-methyleth yl)-, stereoisomer 17.01 0.48
41 (+)-Aromadendren 17.89 1.88
42 (+)-Calarene [(+)-Beta-Gurjunen] 17.99 1.23
45 (+)-Calarene Tetramethyl-Beta-Gurjunen 18.31 6.32
47 Allo-Aromadendren 18.53 0.58
48 Allo-Aromadendren 18.59 6.36
49 Longifolen 18.69 5.70
50 Alpha-Amorphene 18.74 2.98
51 Gamma-Cadinene 18.92 19.47
53 Delta-Cadinene 19.16 0.23
61 Aromadendrene 20.49 0.03
63 Delta-Cadinene 20.86 0.10
64 Hydrocarbons Tetracosamethyl-cyclododecasiloxane 21.00 0.10
65 Gamma-Cadinene 21.16 0.14
66 Alloaromadendrene 21.58 0.05
67 Chlorodecahydroquinoline 21.87 0.69
37 1,4-Methanophthalazine, 1,4,4a,5,6,7,8,8a-octahydro-9,9-dimethyl- 17.54 0.73
7 Di-tert-butyl peroxide 6.15 0.23
13 1, 2, 7, 8-Dibenzocarbazole 10.11 0.04
1 Inorganic compounds Carbon Dioxide 2.34 1.24
2 Carbon Dioxide 2.45 0.07
3 Carbon Dioxide 2.49 0.15
33 Ketones Cyclohexa-2,5-diene-1,4-dione, 2-methyl-5-(4-morpholinyl)- 17.27 0.29

(Continued)

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

TABLE 4 | Continued

Peak Compound RT Concentration

No. (min) (%)

34 2,4(3H,8H)-Pteridinedione, 8-(3,5-Dimethylphenyl)-3-Methyl- 17.33 0.34

35 2-(4’-Chloro)Styrylchromone 2,4(3h,8h)-Pteridinedione, 17.38 0.45
8-(3,5-Dimethylphenyl)-3-Methyl
70 2,5-Di-tert-butylhydroquinone 22.40 0.23
52 Sesquiterpenoids Calamenene 19.00 1.60
54 Alpha-Muurolene 19.22 0.55
55 Alpha-Calacorene 19.32 0.25
59 Tetrahydrosmilagenin 19.88 0.31
60 Alpha-Selinene 20.33 4.01
9 Si-containing compounds Cyclotrisiloxane, hexamethyl- 7.55 0.04
14 Cyclotetrasiloxane, Octamethyl- 10.72 2.78
18 Pentasiloxane, 1,1,3,3,5,5,7,7,9,9-Decamethyl- 12.72 0.02
19 Cyclopentasiloxane, decamethyl- 13.22 3.13
20 Cyclohexasiloxane, Dodecamethyl- 15.65 1.08
21 3,7,7-Trimethyl-11-Methylenespiro(5.5)Undec-2-E 16.13 0.03
25 Cyclotrisiloxane, hexamethyl- 16.58 2.99
26 Oxime-, methoxy-phenyl- 16.63 1.85
27 2,2,4,4,6,6,8,8-Octamethyl-1,3,5,7,2,4,6,8-Tetraoxatetrasilocane 16.74 2.65
28 2-Oxo-4-Nitrosomethyl-6-Trifluoro-Methyl-1,2-Dihydropyrimidine 16.88 0.96
31 Cyclopentasiloxane, Decamethyl- 17.08 1.25
39 Cyclohexasiloxane, Dodecamethyl- 17.78 0.93
40 3-Isopropoxy-1,1,1,7,7,7-hexamethyl-3,5,5 tris (trimethylsiloxy)tetra siloxane 17.84 0.68
56 Dodecamethyl cyclohexasiloxane 19.51 0.14
57 Dodecamethyl cyclohexasiloxane 19.56 0.23
58 Dodecamethyl cyclohexasiloxane 19.65 0.11
68 Hexasiloxane, dodecamethyl 22.02 0.29
69 Hexadecamethyloctasiloxane 22.18 0.08
72 Octa decamethyl, Cyclononasiloxane 23.93 0.03
73 Octadecamethyl Cyclononasiloxane 24.45 0.07
76 Hexadecamethyl Octasiloxane 28.91 0.08
80 Decamethyl cyclopentasiloxane 31.43 0.08

produced by T. harzianum KNU1 significantly enhanced the et al., 2011). Both T. harzianum KNU10 T. reesei KNU4 strains
seedling development and suppressed the radial growth in all showed the same increase in fresh weight (42%). However, the
ginseng root-rot pathogens (Figure 4). More interestingly, VOCs pathogens Trichoderma bioassay exhibited that a mixture of
produced by the T. longibrachiatum strain exhibited a low plant airborne chemicals released from T. harzianum KNU10 strain
growth-promotion effect, however, the T. longibrachiatum-VOCs suppressed the phytopathogens’ growth more effectively. The
significantly suppressed the soil-borne pathogens A. panax, F. airborne chemicals released T. koningii strain which exhibited
oxysporum, and S. sclerotiorum (Table 3). In 2003, Ryu et al. the most effective inhibition against B. cinerea (100% inhibition).
(2003) exhibited that a mixture of airborne chemicals released This result is followed by T. harzianum H22 strain by an
from some bacterial strains of the rhizosphere can promote inhibition percentage of 70%. Changes in these components
the growth of Arabidopsis seedlings. Lee et al. (2016) found because of different physiological activities in vitro largely affect
that tomatoes exposed to airborne chemicals from T. viride the biological functions, e.g., photosynthesis of the plant (Brilli
(BBA 70239) exhibited a larger plant size, a significant increase et al., 2019; El-Maraghy et al., 2020b). The filamentous fungi
in plant biomass (99%), and the development of roots. They isolated from the wheat plant produce volatile constituents that
also observed that the plant growth was dependent on the could promote the growth of various plants and suppress diseases
exposure period to the fungal volatile compounds (Lee et al., (Lyu et al., 2020; El-Maraghy et al., 2021). Subsequently, the
2016). The major volatile component detected from Talaromyces- VOCs-producing fungus Muscodor albus was described to have
VOCs was β-caryophyllene (a terpenoid-like compound) which the capacity for growth improvement and suppression of soil-
significantly promoted the growth of the turnip plant (Yamagiwa borne diseases (Alpha et al., 2015).

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

FIGURE 5 | Phylogenetic analysis of the selected Trichoderma spp. rhizosphere strains; (A) The phylogenic tree showing the lengths of the upright lines are arbitrary;
the lengths of the transversal lines are proportional to genetic distances. Bootstrap frequencies are given for a multiple data set of 100 trials; (B) 18s rRNA gene
sequence similarity for the isolated plant-growth promoting Trichoderma strains.

Headspace GC–MS Analysis of the Fungal and 2-Methyl-propanol formed the major components of the
VOCs volatiles of the plant growth-promoting fungus Phoma sp. GS8-3
The gas chromatography-mass spectrometry analysis (GC–MS) (Naznin et al., 2013). The 2,4-Hexadienal and Benzaldehyde,
of VOCs from the T. harzianum KNU1 strain identified 2,4-bis (trimethylsiloxy) aldehydes were released at 0.07 and
more than 80 different compounds including ketones, 0.56%. 2-Methyl-6-methylene-1,7-octadiene alkene was detected
sesquiterpenoids, and Si-containing compounds. The VOCs of T. as a major volatile component in T. harzianum KNU1-VOCs.
harzianum KNU1 were selected for the GC–MS analysis because Acetonitrile-d3, 2,3-pentadiene, 2,4-dimethyl-, aromadendren,
T. harzianum was the most frequented species during the current dibenzocarbazole, and chlorodecahydroquinoline hydrocarbons
investigation. Besides that, this species achieved adequate were released at low levels from the T. harzianum-VOCs
results in all bioassays. The relative chemical compositions strain, whereas gamma-cadinene was detected as a major
of the main components in T. harzianum KNU1-VOCs were component. Dideuteriohexane 1-pentyl-2,2-d2 acetate was
gamma-cadinene (19.5%), 2-methyl-6-methylene-1,7-octadiene detected as a major ester volatile component. Other esters like
(7.0%), allo-aromadendren (6.4%), (+)-calarene tetramethyl- chlorophenyl 6-(4-morpholinyl) hexyl ether were released but
beta-gurjunen (6.3%), longifolen (5.7%), and alpha-selinene at lower concentrations. Nodulisporium sp. fungus uniquely
(4.0%) (Table 4). Plants and microorganisms comprise many produces many ketone compounds particularly, 1,8 cineole,
volatile natural chemicals. These molecules are chemically 1-butanol, 2-methyl, cyclohexane, propyl, and phenyl ethanol
varied, representing fatty acids, indoles, terpenes, and molecules alcohol, which are the main ingredients of biodiesel when
from other chemical families (Fincheira and Quiroz, 2018). cultured on PDA. Nodulisporium sp. also produces several alkyl
2-Pentenoic acid, 2-ethyl-, methyl ester, and khusilic acids were alcohols, some hydrocarbons, and a few terpenoids (Mends
detected from T. harzianum KNU1-VOCs at a low level of et al., 2012). Alpha-selinene was detected as a sesquiterpenoid
0.03 and 0.15%, respectively. Mhlongo et al. (2018) reported at a relatively high level (4%) and a retention time of 20 min.
that the presence of acids at very low levels in volatiles of Four other sesquiterpenoids, e.g., calamenene and alpha-
the plant growth-promoting rhizomicrobes might reflect the calacorene (naphthalene derivative) were also produced at
redox state of their cells. 1-Butanol, 2,2-dimethyle, and 2- lower levels (Table 4). The endophytic fungus Nodulisporium
amino-1-phenyl-1-propanol alcohols were produced at lower sp. produces cyclohexene, 1-methyl-4-(1-methylethenyl) and
levels than those of acids. Volatiles varied in quantity and 1,4-cyclohexadiene, 1-methyl-, 1–4 pentadiene along with
number according to the incubation period. 3-Methyl-butanol terpenoids and some alcohols of interest as biofuels under

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

microaerophilic growth conditions (Mends et al., 2012). The such as chrysophanol, emodin, and pachybasin, which possess
GC–MS analyses of the VOCs produced by the rhizofungi strong antagonistic properties against pathogenic fungi (Eslahi
produced terpenes including alkenes, alkanes, organic acids, et al., 2021). Ultimately, VOCs produced by Trichoderma spp.
and derivatives of benzene (Naik, 2018). Esters were the most may convey great benefits in the prospective development
effective family of inhibitory compounds tested against fungi, of eco-friendly biopesticides for plant-pathogens control and
for example, 1-butanol 3-methyl-acetate was biologically very proposes the likely use of the natural antimicrobial compounds
effective in reducing the growth of F. solani, S. sclerotiorum, from fungi.
Pythium ultimum, and Rhizoctonia solani (Santra and Banerjee,
2020). Several Si-containing compounds, e.g., oxime, methoxy- CONCLUSIONS
phenyl, were found in T. harzianum strain at low levels,
whereas cyclopentasiloxane decamethyl and cyclotetrasiloxane In this study, the role of volatile compounds produced
octamethyl showed higher emission levels. Etienne and Gefu by fungal strains in plant growth promotion and their
(2009) studied 14 elements including aluminum (Al), boron (B), potential application in biological control were investigated.
calcium (Ca), chlorine (Cl), iron (Fe), copper (Cu), potassium We identified ten rhizosphere Trichoderma strains that showed
(K), manganese (Mn), magnesium (Mg), sodium (Na), sulfur dual antagonistic effects and different morphological features
(S), phosphorus (P), silicon (Si), and zinc (Zn), in two potato (Figure 5). Isolated rhizosphere Trichoderma strains were
cultivars with different fertilization regimes. They found that the investigated for plant growth-promoting traits. The selected
concentrations of Al, Fe, Na, and Si were higher near the ends rhizosphere Trichoderma strains exhibited different capacities
of the tubers and lower at the center of the tuber. The Silicon in their plant growth-promoting traits. Six plant pathogenic
element appeared to be greater near the end of the tuber and fungi were exposed to volatile organic compounds (VOCs)
distributed evenly in the rest of the tuber (Etienne and Gefu, emitted by Trichoderma strains of different species. T. atroviride-
2009). Elements such as Al, Fe, Na, and Si showed a higher VOCs and T. asperellum-VOCs exhibited complete inhibition
concentration at the end and a lower concentration at the center against the key phytopathogen Cylindrocarpon destructans and
of the tuber (Etienne and Gefu, 2009). The potential role of Alternaria panax. Raphanus sativus plant seeds were exposed
d-cadinene in Trichoderma defense needs more investigation to volatile organic compounds (VOCs) emitted by the growing
(Guo et al., 2019). Similar to d-cadinene, caryophyllene was also culture of Trichoderma spp. Exposure to these VOCs emitted
detected only when T. hamatum showed induction to lettuce by Trichoderma strains increased plant biomass by 39.8–
plant growth (Minerdi et al., 2011). Guo et al. (2019) investigated 210% and suppressed the phytopathogens by 16.7–100%. Low
the VOC profiles of three strains of Trichoderma species, levels of Si-containing compounds, e.g., oxime, methoxy-phenyl,
T. hamatum, T. harzianum, T. velutinum, and the common cyclotrisiloxane, hexamethyl, and hexadecamethyl octasiloxane
ectomycorrhizal fungus Laccaria bicolor using the SBSE-GC-MS were found in the VOCs of the T. harzianum KNU1 strain,
technique. They found that cadinene emission was restricted to whereas cyclopentasiloxane decamethyl and cyclotetrasiloxane
the T. harzianum strain. Over 370 secondary metabolites which octamethyl showed higher emission levels. These results suggest
belong to different families of chemical compounds with strong the applicability of Trichoderma-VOCs as biocontrol natural
antagonistic properties have been found to produce Trichoderma products to protect plant crops from soil-borne pathogens and
species (Błaszczyk et al., 2014; Ghorbanpour et al., 2018). The enhance their growth and development.
most important of these secondary compounds are peptaibols
and polyketides (Sood et al., 2020). They are characterized DATA AVAILABILITY STATEMENT
by the presence of amino alcohols and acylated N-terminus
(Zeilinger et al., 2016; Tamandegani et al., 2020). Peptaibol is The original contributions presented in the study are included
produced by the non-ribosomal peptide synthetases (NRPSs) in the article/supplementary material, further inquiries can be
(Sood et al., 2020). The three main gene-encoding NPRSs were directed to the corresponding author.
identified in the Trichoderma genomes (Druzhinina et al., 2011).
Several Trichoderma species were found to produce metabolic
AUTHOR CONTRIBUTIONS
compounds belonging to the group of anthraquinones, pyrones, Both authors listed have made a substantial, direct, and
terpenoids, and epipoly piperazines (Siddiquee, 2014; Zeilinger intellectual contribution to the work and approved it
et al., 2016). The identified terpenoids in Trichoderma include for publication.
tetracyclic diterpenes sesquiterpenes, such as trichothecenes, as
well as triterpene viridin (Zeilinger et al., 2016). T. harzianum, FUNDING
T. koningii, and T. viride species produce the volatile antibiotic
6-phenyl-pyrone, which is responsible for the distinctive coconut This work was supported by the National Research Foundation of
smell (Błaszczyk et al., 2014). Importantly, T. aureoviride, T. Korea (NRF) grant funded by the Korean Government (Ministry
harzianum, and T. viride produce anthraquinone pigments, of Science, ICT, & Future Planning) (NRF-2021R1F1A1063880).

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Joo and Hussein Fungal Plant Growth-Promotion VOCs

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