p-ISSN: 0972-6268

Nature Environment and Pollution Technology (Print copies up to 2016)

2023
An International Quarterly Scientific Journal Vol. 22 No. 3 pp. 1343-1351
e-ISSN: 2395-3454

Original Research Paper https://doi.org/10.46488/NEPT.2023.v22i03.021

Original Research Paper Open Access Journal

Plant Growth Promoting Efficacy of Endophytic Fungi Isolated from the

Terrestrial Plants of North India
Urvasha Patyal* , Vikas Kumar*† , Manoj Singh* and Kulbir Singh**
*Department of Bio-Sciences and Technology, MMEC, Maharishi Markandeshwar (Deemed to be University), Mullana
(Ambala), 133207, Haryana, India
**Department of Civil Engineering, MMEC, Maharishi Markandeshwar (Deemed to be University), Mullana (Ambala),
133207, Haryana, India
†Corresponding author: Vikas Kumar; [email protected]

ABSTRACT
Nat. Env. & Poll. Tech.
Website: www.neptjournal.com Enhanced crop health, which is crucial for sustainable agriculture, is facilitated by a unique
endophyte or endophytic community that is frequently linked to a variety of crops. Plant
Received: 02-12-2022 growth-promoting (PGP) characteristics of endophytes can directly or indirectly boost crop
Revised: 08-01-2023 growth. Endophytic fungi have been proven to create a high percentage of new compounds,
Accepted: 19-01-2023
making them a particularly potential source of physiologically active chemicals. In this study,
Key Words: we have isolated two endophytic isolates, i.e., Paecilomyces sp. (Isolate AT1) and Aspergillus
Endophytes flavus (Isolate AT3), from different host plants, namely Melaleuca citrine and Carica papaya.
Antimicrobial These endophytes have shown significant plant growth-promoting potential toward different
Phylogenetics assays such as IAA production, phosphate solubilization, amylase production, cellulose-
Sustainable agriculture degrading assay, and ammonia production. These endophytic fungi also exhibit visible
Plant pathogens antimicrobial action towards selected crop pathogenic fungi (Aspergillus sp. and Penicillium
sp.). Additionally, these fungal strains are reported for the first time from these plants, as we
have found no reports in the literature. The research aims to explore the growth-promoting
efficacy of endophytic fungi to boost plant growth.

INTRODUCTION Endophytes are a good source of bioactive substances that

improve the nutritional value of the host plant and increase
Endophytes that promote plant growth are symbiotic resistance to pests, diseases, and physical stress (Gouda et al.
organisms (fungi and bacteria) that the host plant uses to 2016). These substances (alkaloids, flavonoids, terpenoids,
protect itself from herbivores in a secretive manner. These are steroids, phenols, phenolic acids, tannins, and peptides),
common microsporic ascomycetes that live inside the healthy which act as enzymes and exhibit antimicrobial and anti-
tissues of living plants for the duration of their lives and are malarial activities, can be used in the food, agricultural, and
found below the layer of epidermal cells in those tissues pharmaceutical industries (Tungmunnithum et al. 2018).
(Mengistu 2020). It offers the plant several advantages, Taxomyces andrenae, an endophytic fungus that belongs to
including the ability to absorb vital nutrients and defense the family of hyphomycetes, produces the anticancer drug
against predatory insects, birds, and animals. Endophytes taxol (Stierle et al. 1993). Plant tissues get inhabited by
are ubiquitous and distinct from all categories of plant PGPE (plant growth-promoting endophytes) and exhibit an
species in an environment, from huge trees to seagrasses. intimate connection within plant tissues, which improves
An estimated 300,000 host plant species were naturally the activities of enzymes and the flow of nutrients, but this
distributed among the endophytic fungi in the temperate much-appropriate hormone distribution encourages plant
region and tropical rainforest (Strobel & Daisy 2003). The development (Hassan 2017). Endophytes have the dynamic
endophytic fungus can create a large range of novel bioactive ability to activate insoluble phosphate and also feed their
secondary metabolites, which are employed in industries to host plant with the right amount of nitrogen (Mehta et
manufacture a variety of natural goods (Sharma et al. 2016). al. 2019).
ORCID details of the authors: In this work, we have selected nine terrestrial plants
Urvasha Patyal: https://orcid.org/0000-0001-5249-0601 of the north Indian regions for the isolation of endophytic
Vikas Kumar: https://orcid.org/0000-0002-6044-3239 fungi. We have isolated several different fungal strains from
1344 Urvasha Patyal et al.

these plants. These strains were restricted to the host plants’ Indole-3-acetic acid (IAA) synthesis test: Fungal cultures
healthy tissues and did not produce any disease symptoms. were dipped into LB broth conical flasks and incubated
When checked for their plant growth-promoting potential, two at 25°C in a BOD shaker for 48 hours. Add 1-2 mL of
out of the nine strains exhibited positive results in different Salkowski’s reagent to the supernatant of the fungal culture
assays. The fungal morphology and molecular identification and keep it in the dark at room temperature for 30 minutes
were determined. For molecular characterization, the DNA of (Bric et al. 1991). A change in the color of the media indicates
these strains was isolated using the CTAB method and then positive results.
subjected to 18S rRNA sequencing by the Sanger dideoxy Amylase activity: Fungal cultures were cut into small
method. The obtained sequence was analyzed, and a consensus discs utilizing a sterilized cork borer and placed over
sequence was prepared, which was then subjected to BLAST the medium plates (Glucose- 0.5g, Yeast extract- 0.05g,
analysis and phylogenetic studies (Kumar et al. 2014, Kumar peptone- 0.25g, agar- 8g, and pH-6) supplemented with 1%
et al. 2017). The consensus sequence was submitted to the soluble starch, and incubated at 25–28°C for 5 days. After
National Center for Biotechnology Information (NCBI). The incubation, cultured plates were flooded with 1% iodine
selected isolates were tested against plant pathogens. In this and 2% potassium iodide (Hankin & Anagnostakis 1975).
assay, some of them showed a positive antimicrobial response. A change in the color of the medium from yellow to pink
Also, these two isolates were further checked for their potential indicates positive results.
to promote plant growth in two different beans, i.e., Vigna
radiata and Vigna mungo. Various growth parameters were Cellulose degrading assay: To select a fungus for
checked for these selected beans using the seed germination cellulolytic activity, fungal isolates were grown in a
assay. It is evident from the results that fungal isolates have carboxymethylcellulose (CMC) agar medium. A disc of 8
promoted plant growth in both selected beans. mm fungal culture was used to inoculate the plates. The test
plates were incubated at 25°C for 5–7 days. To visualize the
MATERIALS AND METHODS hydrolysis zones, the plates were flooded with an aqueous
solution of 0.1% Congo-red (1 mg/mL) for 15 min and
To evaluate the plant growth promotion potential of the washed with 1 M NaCl. Cellulolytic organisms produced a
endophytic fungal isolates, various assays were performed clear zone around the colonies because of the digestion of
to select the potent isolates. The selected fungal isolates were carboxymethylcellulose (CMC) (Dar et al. 2013).
further identified by microscopic characterization as well as Ammonia production activity: Peptone medium (broth)
molecular techniques. was inoculated with different fungal culture discs separately
Collection of plant parts and isolation technique of and incubated at 27°C for 48–72 hours. After incubation,
endophytic fungi: The fresh, healthy, or disease-free plant 0.5 mL of Nessler’s reagent was added individually over
parts were collected from Melaleuca citrina, Phyllanthus the fungal growth in each conical flask (Szilagyi-Zecchinet
emblica, Terminalia arjuna, Eucalyptus globulus, Psidium al. 2014). The change from brown to yellow colour was a
guajava, Azadirachta indica, Acacia nilotica, Tagetes positive test for ammonia production.
erecta, and Carica papaya. These plant parts were carefully
excised with a sterile scalpel, kept inside sterile poly bags, Identification of Endophytic Fungi
and brought to the laboratory for storage at 4°C. All plant Morphological Identification of Fungi: Morphological
parts were cut into small pieces (0.5–1.0 cm) and thoroughly identification of all isolated fungal cultures was done by
washed before processing under running tap water. culturing each culture on PDA medium plates (without
After surface sterilization, potato dextrose agar medium streptomycin) for seven days and continuously observing
supplemented with streptomycin was used to culture the the growth appearance on both sides of the culture plates
fungal isolate and incubated properly at 28±2°C to complete (top and bottom). Based on the standard taxonomic key, we
their growth cycle (Kumar et al. 2016). were able to identify each culture according to its color, the
In-vitro testing for plant growth promotion by endophytic diameter of the fungal colony, texture, morphology, and
isolates: The isolated endophytes were subjected to different dimensions of conidia and hypha (Barnett & Hunter 1998).
plant growth promoting parameters. Microscopic identification: By using the tease-mount
Phosphate solubilizing assay: To check the phosphate method, microscopic slides of each isolated fungal culture
solubilizing capacity, each isolated fungal culture was were prepared using lactophenol cotton blue reagent for
inoculated on the Pikovskayas agar medium plates separately tentative recognition, and each slide was observed under
and incubated at 25-28°C for 48-72 hours (Talukdar a compound microscope. According to the characteristics
&Tayung 2019). of culture, the formation of mycelium and spores helps us

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 PLANT GROWTH PROMOTING EFFICACY OF ENDOPHYTIC FUNGI 1345

identify all unknown isolated endophytic fungi (Aggarwal crops, Vigna radiata and Vigna mungo, the experiment was
& Hasija 1986). first carried out in a growth chamber. Healthy, disease-free
Molecular Identification: Using the 18S rRNA gene seeds were selected and sterilized (Khatun et al. 2008). By
sequencing method, endophytic fungi were molecularly covering the culture with sterile saline containing 0.01% (v/v)
identified. The DNA extraction was carried out by employing Tween (BDH) and spreading the spores with a sterile glass
the cetyl trimethyl ammonium bromide (CTAB) method spreader, it is possible to collect spores from lawn cultures
(O’Donnell et al. 1997). The pure DNA extracted was of the organism on potato dextrose agar media. The spore
amplified by PCR using universal and degenerate primers. solution was then filtered through sterile absorbent cotton
The PCR product was then subjected to gel electrophoresis wool plugs in a row to get rid of any hyphal fragments that
to check the purity of the DNA, followed by analysis might have been present (Jaber & Enkerli 2016). The spore
using Sanger’s dideoxy method to obtain a DNA sequence suspension was then transferred into spray bottles. The sterile
(Cubero 1999). The forward and reverse DNA sequences filter paper was transferred to autoclaved Petri plates, and
of every sample were subjected to the National Centre for seeds of two different samples of crop plants were put in the
Biotechnology Information Basic Local Alignment Search plates in triplicate. The seeds were sprayed with the fungus
Tool (NCBI BLAST) algorithm for identification, and the suspension, and growth conditions of a 12-hour light cycle
phylogenetic tree was also constructed using NCBI BLAST with corresponding 22°C light/ 18°C darkness and 65%
online (https://blast.ncbi.nlm.nih.gov/Blast.cgi). All the moisture content were provided. The seeds were watered
identified 18S rRNA sequences were submitted to GenBank daily, and root and shoot growth were routinely analyzed.
in fast-all format, and accession numbers for the same were
obtained. The identification was confirmed by observing the RESULTS
morphological characteristics through microscopic studies Isolation and Identification of Endophytic Fungi
of the spores, hyphae, and colony.
Various parts were used to isolate fungus strains (stem,
Evaluation for Antagonistic Activity of Isolated root, leaf, bud) of various terrestrial plants (Melaleuca
Endophytic Strains Against Crop Diseases Causing citrina, Phyllanthus emblica, Terminalia arjuna, Eucalyptus
Fungi globulus, Psidium guajava, Azadirachta indica, Acacia
To check the antimicrobial activity of each fungal isolate, test nilotica, Tagetes erecta, Carica Papaya) (Fig. 1). Nine
organisms (Penicillium sp. and Aspergillus sp.) were taken endophytic strains were isolated from all the selected plants.
from the fresh plate and strewn around the fungal culture Identification of Endophytic Fungi
discs in PDA plates and incubated at 25°C for 24-48 hours
(Huang et al. 2000). Antagonism between both the isolated Morphological and microscopic identification: According
endophytic fungi and the fungal pathogen was assessed on an to the standard protocol of (Barnett & Hunter 1998), fungal
individual basis. The testing was carried out in Petri dishes strains were characterized based on their microscopical and
using PDA medium. To remove excess water from the agar cultural properties (Fig. 2). The fungal strains which were
surface, 20 mL of melted medium (40–45°C) was placed into tested positive for different plant growth-promoting activities
sterile Petri plates, cooled, and the plates were left inverted were subjected to molecular identification.
for 24 hours. A mycelial disc of 8 mm in diameter from the Paecilomyces sp. Isolate AT1 was characterized as
actively growing edges of a 4-5 day old culture of a plant endophyte Paecilomyces sp. isolated from the host plant
pathogen was brought adjacent to the media of the Petri plate Melaleuca citrina. The cultural characteristics include fast
to test resistance. An identical 8 mm-diameter mycelial disc growth, powdery, gold, green-gold, yellow-brown, tan
from an actively developing culture of isolated strains (to colored growth. Phialides are bulbous at their bottoms and
be evaluated for antibacterial activity) was placed next to have penicillate crowns. They eventually taper into a long,
the pathogenic fungus after a 24-hour interval. At 25°C, the slender neck. In basipetal continuation from the phialides,
cultures were incubated. Every 24 hours, observations were long, dry chains of single-celled, hyaline to black, smooth
taken to investigate the antimicrobial activity. or rough, round to fusid conidia are formed.

Preparation of the Formulation from Potent Strains Aspergillus flavus Isolate AT3 was characterized as
and Evaluation of Plant Growth Promoting Efficacy endophyte Aspergillus flavus isolated from the host plant
Carica papaya. The macroscopic characteristics include
To confirm the efficacy of seed coating with endophytic the white colony changing to greenish, and the central
fungi to boost the emergence and plant growth of two pulse region showing dark greenish growth. The microscopic

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 1346 Urvasha Patyal et al.

Fig. 1: Endophytic strains isolated from different terrestrial plants.

Identification of Endophytic Fungi

Morphological and microscopic identification: According to the standard protocol of
(Barnett & Hunter 1998), fungal strains were characterized based on their microscopical and
cultural properties (Fig. 2). The fungal strains which were tested positive for different plant
Fig. 1: activities
growth-promoting EndophyticFig. 1:strains
wereEndophyticisolated from
strains isolated
subjected from different
to molecular terrestrial
different terrestrial
identification. plants. plants.

Identification of Endophytic Fungi

Morphological and microscopic identification: According to the standard protocol of
(Barnett & Hunter 1998), fungal strains were characterized based on their microscopical and
cultural properties (Fig. 2). The fungal strains which were tested positive for different plant
growth-promoting activities were subjected to molecular identification.

Fig. 2: Morphology (B, E) (colony appearance) and Microcopy (C, F) (conidia, hypha) view of isolated endophytic strains from different host plants
Fig. 2: Morphology (B, E) (colony(A- appearance) and Microcopy (C, F) (conidia, hypha) view
Melaleuca citrina, D- Carica papaya).

of isolated endophytic
characteristics include a shortstrains fromconidial
and columnar different
head,hostit plants (A- Melaleuca
was determined wereD-
citrina,
that the isolates Carica sp.
Paecilomyces
smooth and brown-colored stripes, biseriatephialides, and (513 bp) and Aspergillus flavus (549 bp). The following
papaya).
globose, smooth conidia. accession numbers represent the fungal pathogens’ gene
sequences that have been submitted to NCBI (Table 1).
Molecular Identification The phylogenetic trees were constructed to represent the
Paecilomyces
Using the 18S rRNA sp.gene
Isolate AT1 was
sequencing characterized
approach, endophytic asevolutionary
endophyterelationship
Paecilomyces sp.organisms
of isolated isolatedwith
fromdifferent
fungi were molecularly identified. Paecilomyces sp. (Isolate biological organisms (Fig. 3).
the host
AT1) plant Melaleuca
and Aspergillus AT3),The
citrina.
flavus (Isolate cultural characteristics include fast growth, powdery,
two endophytic Screening of Endophytic Fungi for Plant Growth
fungi, were amplified and sequenced using universal primers Promoting Activities (PGPA)
gold, green-gold,
Fig.for2:theMorphology
18S rRNA gene.
yellow-brown,
(B, Using
E) (colony tan colored
BLAST appearance)
growth. Phialides are bulbous at their bottoms and
to analyze the and Microcopy (C, F) (conidia, hypha) view
sequence similarity ofcrowns.
have penicillate the resulting
Theysequencing taper Isolated
products,
eventually into aendophytic fungal neck.
long, slender
of isolated endophytic strains from different host plants (A- Melaleuca citrina, D- Carica
culturesInwere screened to
basipetal
Vol. 22, No. 3, 2023 • Nature Environment and Pollution Technology
papaya). This publication is licensed under a Creative
Commons Attribution 4.0 International License
 PLANT GROWTH PROMOTING EFFICACY OF ENDOPHYTIC FUNGI 1347

Table 1: Detail about the fungal isolates with their accession number and GenBank submission name.

S. No. Fungal Fugus Host plant Plant Part Accession number GenBank submission name
1. Paecilomyces sp. (Isolate UP1) Melaleuca citrina Stem OP453360 Paecilomyces sp. Isolate AT1
2. Aspergillus flavus (Isolate UP3) Carica papaya Leaf OP435719 Aspergillus flavus Isolate AT3

Fig. 3: Phylogenetic analysis of endophytic isolates (A) Melaleuca citrine (B) Carica papaya with related genera.

determine the various plant growth-promoting activities exhibited positive antagonistic response against Aspergillus
(Phosphate solubilizing activity, IAA synthesis test, amylase sp. and regarding the other test organism, i.e. Penicillium
Fig. 3: Phylogenetic analysis of endophytic isolates (A) Melaleuca citrine (B) Carica papaya
production activity, cellulolytic activity, HCN production sp, isolate AT3 was somewhere seen inhibiting its growth
activity,with
ammonia production
related genera. activity) and the results are (Fig. 6).
depicted in Table 2. Melaleuca citrine (Isolate AT1) and
Screening
Carica papaya of Endophytic
(AT3) showed highlyFungi forresults
positive Plant inGrowth Promoting
Preparation of theActivities (PGPA)
Formulation from Potent Strains and
different plant growth-promoting assays (Fig. 4). Evaluation of Plant Growth Promoting Efficacy
Isolated endophytic fungal cultures were screened to determine the various plant growth-
Screening of Isolated Fungal Cultures for solubilizing The selected endophytes were screened for their ability to
Antagonistic activity,
promoting activities (Phosphate IAA
colonize two synthesis test,
preferred host amylase
plant species,production
namely Mung bean
Activity
radiata)
activity, cellulolytic activity, HCN production activity, ammonia production activity) andthrough
(Vigna and Urad bean (Vigna Mungo) the seed
Two fungal endophytes (Isolates AT1 and AT3) were inoculation. The fungal elicitors namely Isolate AT1 and AT3
selectedresults
which are depicted
showed highlyin positive
Table 2.plant
Melaleuca
growth-citrine (Isolate
showed AT1)
a positive and induction
growth in plants (AT3)
Carica papaya in terms of the
promoting activities and were then subjected to antimicrobial radicle length and plumule length elongation along with an
showed
assessment highly
against positive
various results in (fungal
test organisms different plant increased
plant growth-promoting
biomass rateassays
in both(Fig.
cases4).
when compared to the
pathogens) (Fig. 5). As a result, isolates AT1 and AT3 control culture. The growth rates were compared with the

Table 2:ofComparison
Table 2: Comparison of PGP
PGP assay of different assaystrains.
endophytic of different endophytic strains.
Isolate Isolate Phosphate
Phosphate solubilizing activity IAA
IAA production Amylase
Amylase production activity Cellulolytic Ammonia
Cellulolytic activity Ammonia production
AT1 positive solubilizing positive
production positiveproduction positive
activity negative
production
AT2 negative activity positive negative
activity negative negative
AT3 positive positive positive positive positive
AT1 positive positive positive positive negative
AT4 negative positive negative negative positive
AT5 AT2
negative negative positive
positive negative
negative negative
positive negative
positive
AT6 negative negative negative positive negative
AT7 AT3
positive positive positive
positive positive
negative positive
negative positive
negative
AT8 positive negative negative negative positive
AT4 negative positive negative negative positive
AT9 positive negative negative negative positive

AT5 negative positive negative positive positive

This publication is licensed under a Creative
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AT6 negative
Commons Attribution 4.0 International License
negative negative positive negative
1348 Urvasha Patyal et al.

Fig. 4: Positive Plant Growth promoting activities by Endophytes isolated from (A) Melaleuca
citrine; (B) Carica papaya.
Screening of Isolated Fungal Cultures for Antagonistic Activity

Two fungal endophytes (Isolates AT1 and AT3) were selected which showed highly positive
plant growth-promoting activities and were then subjected to antimicrobial assessment against
various test organisms (fungal plant pathogens) (Fig. 5). As a result, isolates AT1 and AT3
exhibited positive antagonistic response against Aspergillus sp. and regarding the other test
organism, i.e. Penicillium sp, isolate AT3 was somewhere seen inhibiting its growth (Fig. 6).
Fig. 4: Positive Plant Growth promoting activities by Endophytes isolated from (A) Melaleuca citrine; (B) Carica papaya.
Fig. 4: Positive Plant Growth promoting activities by Endophytes isolated from (A) Melaleuca
citrine; (B) Carica papaya.
Screening of Isolated Fungal Cultures for Antagonistic Activity

Two fungal endophytes (Isolates AT1 and AT3) were selected which showed highly positive
plant growth-promoting activities and were then subjected to antimicrobial assessment against
various test organisms (fungal plant pathogens) (Fig. 5). As a result, isolates AT1 and AT3
exhibited positive antagonistic response against Aspergillus sp. and regarding the other test
Fig. 5: Plant pathogens (A- Aspergillus sp., B-Penicillium sp.).
organism, i.e. Penicillium
Fig. 5: Plant sp, isolate
pathogens (A- AT3 sp.,
Aspergillus wasB-Penicillium
somewhere seen inhibiting its growth (Fig. 6).
sp.).

Fig. 6: Endophytes showing antimicrobial activity against test organisms.

Fig.
help of 6: Endophytes
different graphs. showing
Another antimicrobial
component activity performed
to determine against test organisms.
in triplicates. The seeds of Vigna Radiata and Vigna
Fig. 5: Plant pathogens (A- Aspergillus sp., B-Penicillium sp.).
the seed quality was testified by calculating the seed vigor mungo exhibited a high germination rate when treated with
index.Preparation of the Formulation from Potent Strains endophytic
andisolates AT1 and AT3
Evaluation as compared
of Plant Growthto the control
sample which was treated with water (Fig. 7).
EffectPromoting Efficacy on Mung Bean and Urad
of Culture Supernatant
Bean Germination Effect of Culture Supernatant on Growth of Mung Bean
The selected endophytes were screened for their ability to colonize
(Vigna radiata) two preferred host plant
Plants
A seed germination essay was carried out and seeds germinated
species,
each day namelyforMung
were recorded up to 5bean (Vigna
days. The wasandGrowth
radiata)
experiment Urad promotion
bean (Vigna Mungo)
was studied through
using seed
a 15-day plant growth
inoculation. The fungal elicitors namely Isolate AT1 and AT3 showed a positive growth
Vol. 22, No. 3, 2023 • Nature Environment and Pollution Technology This publication is licensed under a Creative
Commons Attribution 4.0 International License
induction in plants in terms of the radicle length and plumule length elongation along with an
 PLANT GROWTH PROMOTING EFFICACY OF ENDOPHYTIC FUNGI 1349

Effect of Culture Supernatant on Growth of Mung Bean (Vigna radiata) Plants

Fig. 7: Seed Germination percentage essay of Vigna radiata and Vigna mungo.
Fig. 7: Seed Germination percentage essay of Vigna radiata and Vigna mungo.
assay. The Growth
averagepromotion was studied
fresh weight of the using a 15-daywas
seedlings plant growth assay.change
a variable The average
in thefresh weight
lengths of the
of the plumule and radicle
calculated asseedlings
3.612 gwas forcalculated
ControlasC,3.6125.213g for
g Control C, 5.213 as
for Isolate compared
g for to the
Isolate AT1, andcontrol sample
4.055 g for C.AT3.
Isolate
AT1, and 4.055 g for Isolate
The average AT3.ofThe
dry weight theaverage
seedlingsdry
wasweight
calculated as 0.198 g for Control C, 0.213 g for Isolate
of the seedlings was calculated as 0.198 g for Control C, Effect of Culture Supernatant on Growth of Urad
AT1, andAT1,
0.215andg for0.215
IsolategAT3. Bean (Vigna mungo) Plants
0.213 g for Isolate for The seedAT3.
Isolate vigor index
The was calculated to be 1582.05 for Control C, 1840
for isolate
seed vigor index was AT1, and 2811.54
calculated to be for isolate for
1582.05 AT3.Control
Root and shoot
Growthlengths, along with
promotion wasthe average
studied seedling
using a 15-day plant growth
C, 1840 for isolate AT1, and 2811.54 for isolate AT3. Root assay. The average fresh weight
length of treated plants, were recorded and are depicted in Fig. 8. The inoculation of Vigna radiate of the seedlings was
and shoot lengths, along with the average seedling length of calculated as 3.883 g for Control C, 7.210 g for Isolate
seeds
treated plants, werewith endophytes
recorded and areshowed
depicteda variable change
in Fig. 8. The in AT1,
the lengths of the
and 6.148 plumule
g for Isolateand
AT3.radicle
Theasaverage dry weight
inoculation of Vigna radiate
compared to the control sample
seeds with C.
endophytes showed of the seedlings was calculated as 0.197 g for Control C,

Fig. 8: Effect of Supernatent on Root and shoot lengths of Vigna Radiata.

Fig. 8: Effect of Supernatent on Root and shoot lengths of Vigna Radiata.
Effect of Culture Supernatant on Growth ofNature
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Bean (Vignaand
mungo) Plants
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Growth promotion was studied using a 15-day plant growth assay. The average fresh weight of the
1350 Urvasha Patyal et al.

0.208 g for Isolate AT1, and 0.291 g for Isolate AT3. The plant growth by initiating root and shoot length and can be
seed vigor index was calculated to be 1732.5 for Control C, noticed visibly (Fig. 10).
2340 for isolate AT1, and 2392.13 for Isolate AT3. Root
and shoot lengths, along with the average seedling length of DISCUSSION
treated plants, were recorded and are depicted in Fig. 9. The
In this study, according to our research, the isolated fungal
inoculation of Vigna mungo seeds with endophytes showed isolates Paecilomyces sp. and Aspergillus flavus have not
a variable change in the lengths of the plumule and radicle previously been reported in the host plants Melaleuca citrina
as compared to control sample C. and Carica papaya. The plant growth-promoting assays
Upon seed inoculation with fungal elicitors, in comparison of fungal endophytes isolated from host plants Melaleuca
to control it was found that isolates AT1 and AT3 boosted citrina and Carica papaya were analyzed to compare

Fig. 9: Effect of the supernatant on root and shoot lengths of Vigna mungo.

Upon seed inoculation with fungal elicitors, in comparison to control it was found that isolates
AT1 and AT3 boosted plant growth by initiating root and shoot length and can be noticed
visibly (Fig. 10).
Fig. 9: Effect of the supernatant on root and shoot lengths of Vigna mungo.
Fig. 9: Effect of the supernatant on root and shoot lengths of Vigna mungo.

Upon seed inoculation with fungal elicitors, in comparison to control it was found that isolates
AT1 and AT3 boosted plant growth by initiating root and shoot length and can be noticed
visibly (Fig. 10).

Fig. 10: Comparison of growth shown by crop beans A (Vigna radiata), B (Vigna mungo) when inoculated with Control (C) and endophytic fungi
Fig. 10: Comparison of growth shown by(isolates
crop beans
AT1A (Vigna
and AT3). radiata), B (Vigna mungo) when
inoculated with Control (C) and endophytic fungi (isolates AT1 and AT3).
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 PLANT GROWTH PROMOTING EFFICACY OF ENDOPHYTIC FUNGI 1351

and evaluate their effects on root and shoot growth. The Gouda, S., Das, G., Sen, S.K., Shin, H.S. and Patra, J.K. 2016. Endophytes:
isolates’ indirect plant-promoting actions, such as phosphate a treasure house of bioactive compounds of medicinal importance.
Front. Microbiol., 7: 1538. doi: 10.3389/fmicb.2016.01538
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