RESEARCH ARTICLE

Katoch et al., Microbiology 2017;163:840–847

DOI 10.1099/mic.0.000479

An endophytic Fusarium sp. isolated from Monarda citriodora

produces the industrially important plant-like volatile organic
compound hexanal
Meenu Katoch,1,* Kushal Bindu,2 Shipra Phull1 and M. K. Verma2

Abstract
An endophytic fungus, MC_25L, has been isolated from the leaves of MonardacitriodoraCerv. ex Lag., a medicinal and
aromatic herb from the northwestern Himalayas. It produces a fruity fragrance while growing on potato dextrose agar,
suggesting that it is producing volatile organic compounds (VOCs). The endophyte inhibited the growth of plant pathogens
such asSclerotiniasp. and Aspergillusflavus by virtue of VOCs. Identification of MC_25L based on morphological and
microscopic features, as well as ITS-based rDNA sequence analysis, revealed that it is a Fusariumsp. GC–MS analysis
revealed that this endophyte produces a unique array of VOCs, in particular hexanal, p-fluoroanisole, pentafluoropropionic
acid 2-ethylhexyl, (5E)-5-ethyl-2-methyl-5-hepten-3-one, 2-butyl-2-hexanol, (7E)-2-methyl-7-hexadecene and acoradiene.
Three major compounds were hexanal, (5E)-5-ethyl-2-methyl-5-hepten-3-one and acoradiene, and they account for around
84.57 % of the total VOCs. Moreover, of interest was the presence of hexanal, which has applications in the food and
cosmetic industries, as well as in mycofumigation. This is the first report of a fungal endophyte producing the industrially
important plant-like VOC hexanal. Hexanal is also active biologically. Thus this study indicates that Fusariumsp. (MC_25L) is a
potential candidate for the up-scaling of hexanal.

INTRODUCTION among plants, microbes and/or insects, thus influencing the

In the food and cosmetic industries, hexanal is widely used
as a flavouring agent to provide both green character and an
impression of freshness [1]. Nowadays demand for these
natural ‘green note’ products is increasing over synthetic
ones. In 1995, the global flavouring and fragrance market
was estimated to be worth around $20–40 million per year
[2]. In 2013, it reached around $23.9 billion, with an annual
growth rate of 5 –6 %.
Plants are the usual sources for these products. They can
also be produced by enzyme-catalyzed reactions. The for-
mation of these volatiles from hydrolyzed plant oil rich in
linolenic acid requires two enzymes: lipoxygenase and
hydroperoxide lyase. Until now, no microbe was known to
produce hexanal.
Many micro-organisms produce volatile organic compounds
(VOCs) of different chemical classes, such us terpenoids,
straight-chained and branched hydrocarbons, benzene deriva-
tives, naphthalene derivatives, cycloalkanes, alcohols, organic
acids, ketones, and aldehydes for mediating interactions
dynamics of the ecosystem [3–5]. The primary interest in
VOCs comes from their odour and biological properties [6].
Their application and commercialization for flavour and fra-
grance, mycodiesel and mycofumigation started later [7, 8].
This opens up exciting and interesting opportunities in the
field of natural product research, where these molecules may
find use in a multitude of areas.
Endophytes are micro-organisms which reside in the tissues
of almost every plant without causing any detectable symp-
toms [9]. In nature, they help their host to adapt in different
environments by producing various volatile and non-volatile
metabolites [10–12]. Endophytic fungi of several Ascomycota
lineages are capable of producing VOCs [8]. Earlier studies
suggested that sometimes endophytes produce the same bio-
active metabolites that are produced by their host plants [4,
13, 14]. Interestingly, no two organisms produce the same
series of VOCs under identical conditions [15]. Endophytes
exhibit a rich diversity, so chemical/biological novelty is to be
expected [16]. Thus there are many opportunities in the
explortion of endophytes for VOCs.

Received 2 December 2016; Accepted 25 April 2017

Author affiliations: 1Microbial Biotechnology Department, CSIR- Indian Institute of Integrative Medicine, Canal Road, Jammu Tawi, Jammu 180 001,
India; 2Instrumentation Division, CSIR- Indian Institute of Integrative Medicine, Canal Road, Jammu Tawi, Jammu 180 001, India.
*Correspondence: Meenu Katoch, meenusamiksha@rediffmail.com or mkatoch@iiim.ac.in
Keywords: Fusarium sp.; hexanal; VOCs; rDNA; Sclerotinia sp.
Abbreviations: ITS, internal transcribed spacer; VOC, volatile organic compound.
The GenBank/EMBL/DDBJ accession number for the strain MC_25L is KU527796.

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 Katoch et al., Microbiology 2017;163:840–847
Lamiaceae represents the largest plant family containing
aromatic herbages. One of these is Monarda citriodora
Cerv. ex Lag. (lemon bergamot), an annual herbaceous
plant. Traditionally, the plant has been known for its
medicinal properties and fragrance. Tea made from an
infusion of leaves is often used in the treatment of colds,
coughs, fevers and respiratory problems [17]. It is used
as a flavouring agent in the food industry [18]. The
chemical composition of M. citriodora essential oil
extracted from leaves, flowers and dried material has
been studied [19–22]. Its essential oil contains the anti-
septic compound thymol and the insect-repellent citro-
nellol. We chose to explore the endophytes of M.
citriodora because of the medicinal and economic impor-
tance of its essential oil.
In this study, a novel endophyte was reported, which was
isolated from the leaves of this plant, producing a unique
array of VOCs, including an industrially important one –
hexanal. It was also evaluated for biocontrol/
mycofumigation potential.
METHODS
Isolation and characterization of MC_25L
Endophyte MC_25L was isolated from the leaves of M. cit-
riodora collected during March–April 2013 from the Shiva-
lik hills of Jammu and Kashmir (32.73 N 74.87 E) as
described earlier by Strobel and Daisy [23], with slight mod-
ifications. The culture was submitted to the Sir R N Chopra
Microbial Resource Centre, Jammu, India with accession
no. MRCJ-512. The fungus, by virtue of a simple smell test,
yielded a fruity odour. The endophyte was identified on the
basis of the morphophological and microscopical character-
istics and by internal transcribed spacer (ITS)-based rDNA
sequencing. Genomic DNA of the endophytes was extracted
from the in vitro grown biomass of the endophyte [24]. The
ITS region (ITS1-5.8S-ITS2) was amplified and sequenced
with the universal primers, with a previously described reac-
tion mixture and programme [25, 26]. The resultant
sequence (KU527796) was submitted to GenBank and ana-
lysed using the NCBI BLASTN tool for final identification
[27]. For the phylogenetic tree, the endophytic ITS sequence
and the downloaded sequences of their nearest neighbours
were evaluated using MEGA4 software [28].
Bioactivity of MC_25L
Fungal pathogens used and antagonistic studies
Antagonistic activity of MC_25L was evaluated against com-
mon phytopathogens viz. Aspergillus flavus (accession num-
ber MTCC 1783), Fusarium solani (MTCC 350), Sclerotinia
sp. (MTCC 7114), Colletotrichum capsici (MTCC 2071) and
Aspergillus fumigates (MTCC 343), procured from the
Microbial Type Culture Collection and Gene Bank (MTCC),
India. The antagonistic potential of the MC_25L was
assessed through co-culture assay, culture filtrate assay and
fumigation assay [29, 30]. All the experiments were per-
formed in triplicate and mean values were calculated.
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Co-culture assay
Co-culture assay was used to evaluate MC_25L for its ability
to inhibit the fungal pathogen’s growth on potato dextrose
agar (PDA) [29]. Petriplates containing 15–20 ml PDA were
prepared. Bits of MC_25L and pathogen (0.5 cm each) were
co-cultured at the two opposite ends of the plates. The plates
were incubated at 25±2  C after being sealed with parafilm.
The pathogens alone without MC_25L served as the control.
After 7 days, the diameter of the pathogen’s growth in the
presence/absence of MC_25L was measured. The percentage
antagonism was calculated using the formula A (%)=
[(CDC CDT)/CDC]*100, where CDC represents the diame-
ter of the pathogen’s growth (measured in mm) in the absence
of MC_25L, and CDT is the diameter of the pathogen’s
growth in the presence of MC_25L [31, 32].
Culture filtrate assay
MC_25L was cultured in 50 ml of potato dextrose broth
(PDB) in 250 ml Erlenmeyer flasks by inoculating two plugs
(0.5 cm) of actively growing endophytic fungus. The flasks
were incubated for 10 days in a rotatory shaker (150 r.p.m.)
at 25±2  C. The cultures were centrifuged to remove bio-
mass so as to collect broth containing metabolites [33].
MC_25L culture filtrate (200 µl) was spread on PDA plates
to test its antagonistic activity. On drying of the filtrate,
respective pathogenic fungus (0.5 cm plug) was inoculated
in the centre of the PDA plate. Simultaneously, PDA plates
containing pathogenic fungi without MC_25L culture fil-
trate served as the control. The plates were incubated at 25
±2  C. After 7 days, the diameter of the pathogen’s growth
in the presence/absence of MC_25L culture filtrate was
monitored and the percentage antagonism was calculated.
Fumigation assay
The assay was conducted in PDA Petri plates, which were
inoculated separately with MC_25L and pathogenic cultures
(0.5 cm plug from actively growing fungus). The plates were
incubated at 25±2  C after being sealed with parafilm. After
2–4 days, the lids were removed and the Petri dish inocu-
lated with MC_25L culture was inverted on the Petri dish
inoculated with pathogen under aseptic conditions, and
then the dishes were tightened with a double layer of
parafilm. The plates were again incubated at 25±2  C for
7 days. the Petri dish containing PDA without MC_25L
inverted on the Petri dish inoculated with the pathogen
served as the control. The diameter of the pathogen’s
growth in the presence/absence of MC_25L was monitored
and the percentage antagonism was calculated.
Cytotoxic and antimicrobial studies
The DCM extract of MC_25L was also evaluated for its
cytotoxic and antimicrobial effects against different
human cancer cell lines (HCT-116, A549, PC-3 and
T47D) and pathogens using MTT and microdilution
assay, respectively [26].
Fermentation and extraction
For the extraction of VOCs, the endophyte MC_25L was
cultured on a set of five PDA plates for a period of 15 days
 Katoch et al., Microbiology 2017;163:840–847

at 25±2  C in an incubator (New Brunswick, USA). One RESULTS AND DISCUSSION

block (0.5 mm) of 10-day-old fungal growth was used as
inoculum. After 15 days, the fungal growth of the five Petri
dishes was homogenized thoroughly with 12.5 ml of metha-
nol. Homogenate was extracted with one volume of methy-
lene chloride (DCM; HPLC grade) [34]. The extraction
process was repeated four times. For the control, a set of
five PDA plates incubated at 25±2  C for 15 days were
extracted in a similar way.
GC–MS analysis of fungal VOCs
Volatile components were analysed in the control and the
MC_25L endophytic extract using a Varian mass spectrom-
eter 4000 coupled with a GC 3800 equipped with a CP-Sil-8
capillary column (30 m0.32 mm0.25 µm film thickness).
The carrier gas was helium, with a flow rate of 1 ml min 1.
The column split ratio was 1 : 150. The temperature pro-
gramming for the column oven was 60  C for 5 min; from
60  C to 250  C at a 3  C min 1 rising rate; hold for 7 min.
For GC–MS detection, an electron ionization system was
used with an ionization energy of 70 eV. Mass spectra were
recorded over a 50–300 amu range at one scan per second.
The extract was prepared as 10 mg ml 1 in ethyl acetate and
2 µl of the sample was injected automatically in the split
mode. The injector and detector temperatures were set at
280  C.
MC_25L as an endophyte
Endophyte MC_25L was isolated from the leaves of the plant
host M. citriodora. The colonies on PDA were 80–85 mm after
10 days at 25  C. Initially they were white and later started to
convert to a peach colour from the centre to periphery. There
was profuse aerial mycelium, the margins were regular and it
was reverse concolourous. Microscopically, the fungus pro-
duced single-celled straight, ovoid and hyaline micro-conidia.
Collectively, these morphological/microscopical features
strongly support the placement of the present isolate as a spe-
cies of Fusarium sp. (Fig. 1). While growing on PDA, endo-
phyte MC_25L produced a fruity fragrance.
Phylogenetic position of MC_25L
Furthermore, ITS sequence data showed that the endophyte
is a strain of the genus Fusarium (Fig. 2). The ITS1 5.8S
ITS2 region of the ribosomal gene of MC_25L showed a
maximum homology of 100 % with Fusarium sp. strain
C_1_BESC_294z, Fusarium redolens, Hypocreales sp. SN-
2008 and 99 % sequence homology with Fusarium acumi-
natum isolate 30SS3, as presented in the phylogenetic tree
(Fig. 2). The phylogenetic tree contained two clusters. Taxo-
nomically, MC_25L is placed in a separate cluster to the one
containing Fusarium oxysporum strains.

Initial identification of the VOCs produced by the endo- Bioactivity of MC_25L

phyte MC_25L was made via library comparison using the
National Institute of Standards and Technology (NIST)
database, and thus all the chemical compounds described in
this report use the NIST database chemical terminology.
GC–MS analyses were performed three times and mean val-
ues were presented.
The VOCs of MC_25L exhibited varying degrees of antagonis-
tic activity against the tested pathogens (Table 1). This ranged
between 24 and 52 %. The VOCs of MC_25L showed mycelial
(a)

Quantification of hexanal
GC–MS analysis was carried out on an Agilent GC–MS
5975 C fitted with an HP-5 ms fused silica capillary col-
umn (30 m0.25 mm id, film thickness 0.25 µm). The
temperature programming of the oven was: from 50 to
250  C at a 5  C min 1 rising rate; then up to 280  C; then
hold for 2 min. Helium was used as the carrier gas, with a
flow rate of 1 ml min 1. Mass spectra were recorded over (b)
a 30–400 amu range at one scan per second with EI at
70 eV.
Identification of the hexanal was made by comparing the
mass spectra with those of the internal reference mass spec-
tra library (Wiley/NIST/Pfleger/) or with authentic com-
pound [procured from TCI chemicals (India) Pvt. Ltd], and
was confirmed by comparison of the retention indices with
those of authentic compounds or with those reported in the 50.0 µm

literature database.
Quantification of hexanal was carried using the external Fig. 1. (a) A colony morphology of MC_25L, an endophyte, associated
standard method. Excellent calibration curves were obtained in nature with Monarda citriodora, identified as Fusarium sp. growing
for hexanal in the concentration range 50.0–800.0 µg ml 1 on PDA. The fungus was found to produce a fruity fragrance. (b) Light
(R2 >0.999). The LOD and LOQ of hexanal were 9.9 and microscopic view of MC_25L at 400x.
38.8 µg ml 1, respectively.
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22 Hypocreales sp. EU754933

13 Fusarium acuminatum JX114782

0.02 99 Fusarium sp. KT347158

MC 25L
93
Fusarium redolens HQ703404
99
Fusarium sp. KJ567458
Fusarium oxysporum KF998987
100 Fusarium oxysporum KF264963
Fusarium solani FJ426390
Colletotrichum boninense JQ676184
Colletotrichum siamense KP748202
100
Colletotrichum gloeosporioides KM520010
97
76 Colletotrichum fructicola KP748219

Fig. 2. Phylogenetic position of MC_25L. The evolutionary history was inferred using the neighbour-joining method. The percentage of
replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) is shown next to the branches.
The evolutionary distances were computed using the maximum-composite-likelihood method. Phylogenetic analyses were conducted
in MEGA4 [28].

1
growth inhibition of 52 % against Sclerotinia sp., followed by found for human microbial pathogens up to a 100 µg ml
50 % against A. flavus, 36.6 % against F. solani and 24.44 % concentration.
against A. fumigatus. They showed no growth inhibition
against C. capsici. These results suggest that the VOCs of
VOC production by MC_25L
MC_25L are not only active, but specific as well. The fungus, by virtue of a simple smell test, yielded a fruity
fragrance, indicating the production of VOCs. That the VOCs
Further, the culture filtrate assay displayed better activity of MC_25L possessed antagonistic activity against plant
(65 %) against Sclerotinia sp., suggesting that compounds pathogens was subjected to further study. For qualitative anal-
other than VOCs are also contributing towards the antago- ysis of the VOCs produced by this organism, it was grown on
nistic activity against Sclerotinia sp. This result suggested PDA and PDB for 10 days. Headspace analyses of solid and
that antagonistic metabolites are produced in greater quan- liquid culture of MC_25L were carried out. The DCM extract
tity. In contrast, the co-culture assay did not show any of MC_25L was also analysed through GC–MS and com-
antagonistic activity (50 %). This might be because pounds were identified on the basis of retention time and
MC_25L is producing molecules other than antagonistic peak mass compared with the NIST database information
ones or not diffusing through agar. (Table 2). A high match score value supports the results. This
The extract of MC_25L was also active against different endophyte produces a unique array of VOCs, in particular
human cancer lines HCT-116, A549, PC-3 and T47D,with hexanal, p-fluoroanisole, pentafluoropropionic acid 2-ethyl-
IC50 values 10–12.7 µg ml 1. However, no inhibition was hexyl, (5E)-5-ethyl-2-methyl-5-hepten-3-one, 2-butyl-2-hexa-
nol, (7E)-2-methyl-7-hexadecene and acoradiene (Fig. 3 and
Table 2). From GC–MS analysis, the relative percentage area
Table 1. The biological activity, represented as percentage antagonism or was calculated for each compound. Hexanal (RT 7.215 min)
growth inhibition, of MC_25L against a panel of fungal phytopathogens constituted about 43.9 % of the total VOCs, followed by (5E)-
Pathogen Antagonism percentage*
5-ethyl-2-methyl-5-hepten-3-one (RT 22.687, 15.9 %) and
acoradiene (RT 46.988 min, 24.6 %). Collectively, the aldehyde
Co-culture Culture filtrate VOCs present in the DCM extract of MC_25L comprised the great-
Fusarium solani 31.2 46.66 36.6
est percentage of total VOCs, followed by terpene and ketone.
Sclerotinia sp. – 65.21 50
Hexanal was confirmed and quantified (7.8 mg l 1) using its
standard (Figs 4, 5).
Colletotrichum capsici – 47.36 0
Aspergillus flavus 28.6 50 52
Aspergillus fumigatus 29.4 11 24.44
DISCUSSION
Fusarium sp. was found in this study to be associated with
*Antagonism values were calculated as the percentage of lateral growth
leaves of M. citriodora as an endophyte. It was identified
inhibition relative to the control test organism. The values are an average
of three similar observations. morphologically, microscopically and through ITS-based
rDNA sequence analysis. The VOCs of MC_25L showed
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kCounts TIC 25L(P) 10-23-2013 9-45-56 PM. SMS 4000 Centroid

lonizat... 40:500

HEXANAL

(7E)-2-Methyl-7-hexadecene
7.215 min
50

46.988 min
Acoradiene
40

2-Butyl-2-hexanol
23.908 min
p-Fluoroanisole
30

13.729 min

44.946 min
20

10

10 20 30 40 50 60 70 minutes
Seg 1 Seg 2, <No Description, Time: 4.00-77.00
655 1338 2021 2704 3388 4072 4755 Scans

Fig. 3. GC–MS chromatogram showing secondary metabolites produced by MC_25L.

varying degrees of antagonistic activity, being highly active (5E)-5-ethyl-2-methyl-5-hepten-3-one, 2-butyl-2-hexanol,

against Sclerotinia sp. and A. flavus (50 %), and moderately (7E)-2-methyl-7-hexadecene and acoradiene. In this study,
or weakly active against F. solani (36.6 %) and A. fumigatus the VOCs for this fungal isolate followed a distinct pattern,
(24.44 %). They showed no growth inhibition against since no other Fusarium isolate that has been studied has
C. capsici. It is well documented that in nature these gaseous revealed an identical pattern to this one. Hexanal (RT
substances might be involved in inhibiting or killing com- 7.215 min) constituted about 43.9 % of the total VOCs in
peting micro-organisms, either specifically or nonspecifi- the DCM extract of MC_25L. Hexanal is also known as hex-
cally [35]. Thus the VOCs of this organism can be exploited analdehyde or caproaldehyde. Similar to the present study,
to inhibit specific pathogens of agricultural crops, and can hexanal has been reported as one of the VOCs from virgin
thus be used in disease management. coconut oil, potato tubers inoculated with Phytophthora
GC–MS analysis of MC_25L extract revealed the presence infestans or Fusarium coeruleum and three head blight caus-
of a unique array of volatile compounds, particular hexanal, ing Fusarium graminearum chemotypes [36–38]. Hexanal is
p-fluoroanisole, pentafluoropropionic acid 2-ethylhexyl, an alkyl aldehyde with or without double bonds, and is a

Fig. 4. (a). GC–MS chromatogram showing hexanal. (b) MS scan of hexanal.

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Table 2. Volatile organic compounds (VOCs) produced by Fusarium sp., as determined by GC–MS

RT (min) Compound Area Amount (%) Score match Molecular weight

(kCounts)

7.215 Hexanal 37 935 43.974 95.59 100

13.729 p-fluoroanisole 4 610 5.344 94.37 126
21.716 Pentafluoropropionic acid, 2-ethylhexyl 2 658 3.081 88 276
22.687 (5E)-5-ethyl-2-methyl-5-hepten-3-one 13 761 15.952 80 154
23.908 2-butyl-2-hexanol 4 665 5.408 91 158
44.946 (7E)-2-methyl-7-hexadecene 1 378 1.597 85 238
46.988 Acoradiene 21 260 24.645 81 204

dominant compound released by plant material through the
lipoxygenase pathway after tissue disruption; it is used in
the food and cosmetic industries to produce fruity flavours
[39]. Furthermore, it has been approved as a food additive
by the US Food and Drug Administration (FDA) [40]. The
VOCs of MC_25L showed antagonistic activity against
major phytopathogens, which might be attributed to the
presence of hexanal. Earlier studies also reported that its
vapours suppress spore germination, mycelia growth and
the fungal-derived cell-wall-degrading enzymes of posthar-
vest pathogens [40–43]. Using hexanal as a fungicide
presents lower human risks and animal toxicity, and causes
less environmental pollution, while the research and devel-
opment costs are lower than those for synthetic fungicides
[44]. In industry, it is produced through either chemical
processes or biotechnological interventions [45, 46]. Until
now, no endophyte was known to produce hexanal (associ-
ated with fruity flavour and antagonistic potential) as part
of its VOCs. Another product of interest is 2-butyl-2-hexa-
nol. It is an alcohol that is also used in the flavouring indus-
try to produce fruity flavours. It is also a ‘green note’.
Furthermore, acoradiene was present in the fungal VOCs as
a second major VOC (24.6 %). It is also a major VOC in the
volatiles of F. oxysporum isolate 21, Fusarium sambucinum
and Phomopsis sp. [47–49]. The volatiles of F. oxysporum
Hexanal
Response
1.00e+005
5.00e+004
showed strong mortality to Meloidogyne incognita, a
nematode. No later report on its specific activity against
nematodes was found in the literature [50]. Sanchez-Ortiz
et al. [51] reported that this compound is also responsible
for the antifungal and anti-oomycete activities. Fiers et al.
[52] also reported that the VOCs of F. culmorum, including
acoradiene, decreased the growth of Cochliobolus sativus by
13 to 17 % in barley. Singh et al. [49] reported its involve-
ment in antagonistic action, and its biofuel potential.
(7E)-2-methyl-7-hexadecene, a third major component of
the present study, is also reported to be a VOC of Psoralea
carylifolia L. (Cullen corylifolia). Other products of interest
include ketone [(5E)-5-ethyl-2-methyl-5-hepten-3-one] and
acid (pentafluoropropionic acid, 2-ethylhexyl) [53]. Pro-
pionic acid, which contributes towards biological activity
against plant pathogens, was reported from Nodulisporium
sp. [54].
Another volatile compound was found in the DCM extract
of MC_25L: p-fluoroanisole. It is widely used in the fertil-
izer, chemical and agricultural industries. Anisole or
methoxy benzene is a colourless liquid with a smell reminis-
cent of aniseed, and in fact many of its derivatives are found
in natural and artificial fragrances. 2,4,6-trichloroanisole
(TCA) is a chemical compound that is a chlorinated deriva-
tive of anisole. TCA is a fungal metabolite of 2,4,6-trichloro-
phenol, which is used as a fungicide.
In conclusion, in this study MC_25L produces a unique
range of industrially important VOCs that are also bioac-
tive. Although MC_25L (Fusarium sp.) is genetically (ITS
region) similar, it has the ability to produce VOCs that are
different from those of other Fusarium spp. This is the first
report of a fungal endophyte producing the industrially
important plant-like VOC hexanal. Thus this study indi-
cates that MC_25L is a potential candidate for the up-
scaling of hexanal.

0 Funding information
0 500 The research conducted in this paper was supported by the DBT spon-
sored project GAP1182 (BT/PR4669/PBD/17/784/2012), Government
Concentration (µg ml–1) of India.

Acknowledgements
Fig. 5. Calibration curve of hexanal. This article bears the Institutional Publication No. IIIM/2032/2017
GAP1182. We acknowledge the DBT, Govt of India, for financial

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assistance and the Council of Scientific and Industrial Research (CSIR)
for providing the research platform.
Conflicts of interest
The authors declare that there are no conflicts of interest.
Ethical statement
This study does not include experiments on animals or humans.
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