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Homeopathic medicines as new strategy against plant pathogenic fungi

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DOI: 10.54085/ap.2023.12.1.19

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Annals of Phytomedicine 12(1): 1-15, 2023

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Original Article : Open Access

Homeopathic medicines as new strategy against plant pathogenic fungi

Dheer Kumar, Samir K. Biswas , Shivam Kumar, Girijesh Kumar Jaisval, Deepoo Singh, Ravi Kumar, Rahul Kumar Tiwari*,
Milan Kumar Lal* and Ravinder Kumar*
Department of Plant Pathology, Chandra Shekhar Azad University of Agriculture and Technology, Kanpur, 208002, Uttar Pradesh, India
* ICAR-Central Potato Research Institute, Shimla, 171001, Himanchal Pradesh, India

Article Info Abstract

Ar tic le histo ry Nowadays, homeopathy is being rapidly practiced almost all over the world for preventing or curing
Received 10 January 2023 human and plant diseases. In the current study, homeopathic medicine, i.e., Arnica montana, Thuja
Revised 5 March 2023 occidentalis, sulfur, and silicea were applied against plant disease causing pathogens, i.e., Fusarium
Accepted 6 March 2023 oxysporum f. sp. lycopersici, Ascochyta rabiei, Alternaria solani, Septoria lycopersici, and Phomopsis
Published Online 30 June-2023 vexans. The radial mycelium growth of these species was found to be significantly decreased with the
increased concentrations of homeopathic medicine under in vitro condition. At 8 days after inoculation,
Ke ywor ds
Homeopathic medicine the highest inhibition of mycelial growth of F. oxysporum f. sp. lycopersici, A. solani, S. lycopersici, A.
Antifungal activities rabiei, and P. vexans over control was observed with T. occidental at 200 ppm (73.73%), T. occidentalis
Induce resistance at 200 ppm (76.73%), A. montana at 200 ppm (64.22%), silicea at 200 ppm (59.77%), and sulfur at 200
Plant pathogenic fungi ppm (53.82%), respectively. Accordingly, these fungal species minimal radial growth was as follows:
22.80 mm, 18.10 mm, 31.66 mm, 34.91 mm, and 38.12 mm. Such findings outline the potential of
homeopathic medicine treatment in the reduction of mycelial growth of several fungal species affecting
consumed crops.

1. Introduction in bean plants, growers turned to homoeopathic remedies containing

phosphorus and Calcarea carbonica (Rissato et al., 2016). Recently,
Homeopathy simply means treating diseases with remedies, homeopathic medicine, e.g., A. montana, T. occidentalis, sulfur, and
prescribed in minute doses. It is based on the natural law of healing silicea were applied against plant disease-causing pathogens, i.e., F.
“similar similibus curantur”, which means “likes are cured by likes” oxysporum f. sp. lycopersici, A. solani, S. lycopersici, A.rabiei, and
and was given a scientific basis in the early 19th century. Homeopathy P. vexans. A. montana is a beautiful mountain daisy mostly found in
postulates that suppressive activities, that go against the vital acidic soils and mountain pastures. It exhibits anti-inflammatory,
principle, are to blame for the onset of disease in living beings (Bellavite antiseptic, antifungal, and antibacterial activities (Conforti et al.,
et al., 2007). Nowadays, homeopathy is being rapidly practiced 1997). Hanif and Dawar (2015), reported that the mycelial growth
almost all over the world. In India, it has become a household name of F. oxysporum, Rhizoctonia solani, and Macrophomina phaseolina
due to its safety and the gentleness of its cure. It has been stated that was significantly inhibited by A. montana. T. occidentalis is an
about 10 per cent of the Indian population slowly depends on evergreen coniferous tree used intensively in the homeopathic system
homeopathy for healthcare needs which are considered the second of medicine (Hulten and Mistry, 1986; Alam et al., 2022). It possesses
most popular system of medicine in the country. Nearly 70 per cent several medicinal values including antiviral, antifungal, antidiarrheal,
of homeopathy medicines are derived from plant parts, i.e., fruit, and antioxidant properties (Caruntu et al., 2020). Khanna and
stem, bark, flower, leaf, stigma, or root, as well as a non-woody Chandra (1989), obtained positive results in the control of tomato
plant and some other herbs. Most homeopathic medicines are (Solanum lycopersicum L.) rot caused by F. roseum, after the
prepared from natural herbs. Generally, they are non-hazardous and application of T. occidentalis. To combat soil-borne infections,
eco-friendly target-specific modes of action with least side effects, researchers have experimented a variety of formulations, including
which prevent or cure human diseases. Diseases in plants are caused bacterial suspensions, fungal spores, and powdery preparations of
by an imbalance of vital energy, which can also lead to the plants fungal mycelium. The population of soil-borne plant pathogens and
demise if their production is drastically reduced (Nazarov et al., plant parasitic nematodes has been reduced in several experiments
2020). However, homoeopathic medicine helps to mitigating negative where microbial antagonist was added to the soil (Topalobic et al.,
effects on vital energy and restore equilibrium by boosting the plant’s 2020). Root rot pathogen colonization can be reduced and plant
defensive mechanism (Biswas et al., 2002). To eliminate white mold growth can be enhanced using microbial antagonists such as
Trichoderma spp., Paecilomyces lilacinus, Verticillium
Cor respo nding autho r: Dr. Samir K umar Biswas
Professor Head, Department of Plant Pathology, Chandra Shekhar
chlamydosporium, Bacillus spp., Stachybotrys atra, Pseudomonas
Azad University of Agriculture and Technology, Kanpur-208002, Uttar aeruginosa, and Gliocladium virens (Brotman, 2013; Loser et al.,
Pradesh, India 2021). By releasing chitinolytic enzymes and generating
E-mail: samir krbiswas@ rediffmail.c om mycoparasitism inhibitory chemicals, antagonistic bacteria can
Tel.: +91-9452522504 regulate soil-borne diseases (Sharp et al., 2013). Therefore,
Copyright © 2023 Ukaaz Publications. All rights reserved.
Email: [email protected]; Website: www.ukaazpublications.com
2

mycoparasitism inhibitors may be responsible for the suppression 2. Materials and Methods
of soil-borne diseases (Lahlali et al., 2022). Chitinase enzyme is
responsible for the breakdown of b-glucan, chitin, and 2.1 Experimental site
polysaccharides in fungal cells, hence killing the pathogen (Veliz et All experiments were conducted in the laboratory of Department of
al., 2017). Rex et al. (2019), reported that the antifungal activity of Plant Pathology, C.S.A., University of Agriculture and Technology,
crude plant extracts of ten plant species, viz., Neem (Azadiracta Kanpur during the 2020-2022 periods.
indica), Prosopis (Prosopis juliflora), Onion (Allium cepa),
Eucalyptus (Eucalyptus obliqua), Pungam (Pongamia pinnata), Curry 2.2 Isolation and purification of the pathogen
leaf (Murrya koenigii), Garlic (Allium sativum), Henna (Lawsonia F. oxysporum f. sp. lycopersici, A. rabiei, A. solani, S. lycopersici,
inermis), Turmeric (Curcuma longa) and Ginger (Zingiber officinale) and P. vexans were isolated from soil and infected plant parts, i.e.,
were evaluated against A. solani (Ellis and Martin) by using poisoned leaves, root, stem, and fruit. The selected samples were rinsed in tap
food technique under in vitro conditions and found that turmeric
water to remove dust particles, and surface contaminants.
extract recorded minimum radial mycelial growth of 9.50 mm with
Subsequently, the young diseased parts of the leaves, stems, roots,
maximum per cent inhibition of 89.44 over control, followed by
and fruits were cut into small bits along with some healthy portions
garlic observed (12.80 mm, 85.77 per cent) and eucalyptus (24.27
using a sterilized blade. The cut pieces were then dipped and sterilized
mm, 73.03 per cent), respectively. Volatile organic compound (VOC)
with 0.1% sodium hypochlorite solution for 20-30 sec using sterilized
from plants known to have varied potential sources for antimicrobial
forceps. They were then thoroughly rinsed thrice in sterilized water
activity against soil-borne pathogens and evaluate in vitro antifungal
to remove any remaining trace of sodium hypochlorite solution. The
activity of volatile organic compounds of carvone and citronellol
excessive moisture was removed by placing samples in the fold of
against spore germination of F. oxysporum f.sp. lycopercisi (Praveen
pre-sterilized blotting paper. Petri-plates were sterilized at 160-180
et al.,2021). Akshaya et al. (2021), reported that the ethyl acetate o
C for 2 h in a hot air oven. The surface of the pieces was transferred
and methanolic fractions of cell-free culture filtrate (18 days old) of
in petri-dishes using sterilized forceps. Petri-dishes used in the
O. sinensis at a concentration of 3000 ppm showed maximum
experiment were previously poured with sterilized 20 ml PDA
inhibition of mycelial growth of Fusarium oxysporum f. sp.
medium under laminar flow. The surface sterilization of three to four
lycopersici (43.10 and 46.6 per cent, respectively), followed by
pieces of the diseased part of tomato leaves and roots, chickpea
Fusarium oxysporum f. sp. cubense (42.20 and 41.10 per cent,
stem and root, and brinjal fruit were placed in each petri-dish at an
respectively), when tested by poisoned food technique. Gayathiri et
equal distance fields (Synder and Hansen, 1940; Dhingra and Sinclair,
al. (2021), reported the standard samples of papaverine tested against
1995). These petri-dishes were properly marked with glass marking
mycelial growth and conidial germination of C. gloeosporioides
exhibited 100 per cent inhibition of mycelial growth of C. pencils indicating the date of isolation. They were finally sealed
gloeosporioides at 1500 ppm and 3000 ppm in agar well diffusion with parafilm tape, incubated at 25-28oC in a BOD incubator, and
assay and paper disc assay which confirmed the presence of observed periodically for fungal growth on 24 h intervals. Once the
antimicrobial metabolites from culture filtrates of G. lucidum. Maurya mycelial growth occurred, the hyphal tips for the advancing mycelium
et al. (2021), was carried out to check inhibitory effect of botanicals were transferred aseptically into the sterilized culture tubes containing
against Ustilaginoidea virens. Among botanicals, neem oil at both 2% potato dextrose agar (PDA) medium for further purification,
concentration, i.e., 5 per cent and 10 per cent was found highly identification, and maintenance of culture. The purification of fungi
effective and inhibit the growth by 66.97% and 70.14%, respectively isolates was processed by adopting a single spore isolate technique
followed by onion (62.89%, 67.64%), garlic (58.82%, 64.25%), ginger (Choi et al.,1999).
(55.88%, 63.57%) and tulsi (46.40%, 54.98%). Hence, plant roots 2.3 Identification of the pathogen
can be protected from soil-borne pathogens via the barrier created
by the microbial antagonists. The use of homeopathic medicine to The pathogen was identified based on its morphological and cultural
control various pathogens affecting fruits was found highly effective characteristics, as well as its pathogenic behavior toward the host.
and environmentally safe. For instance, the inhibitory effects of T. The A. solani stands distinct from other genera by its transversely
occidentalis against Alternaria alternata, Fusarium moniliforme, and longitudinally septate (muriform) conidia. The conidia show a
Gloeosporium psidii, Colletotrichum gloeospoioides, Pestalotia spp., distinct beak which may be short or very long. The conidia are
and other fruit rot pathogens have been previously reported (Chandra usually formed in chains, they were in the current case dark brown
et al., 1981; Wilson et al., 1991; Khanna et al., 1992; Baviskar et al., and typically obclavate (Alhussaen, 2012). The identification of F.
2015). Arie (2019), reported that sulfur is a very effective oxysporus f. sp. lycopersici was possible from its hyphae which are
homeopathic medicine against some plant pathogenic fungi such as hyaline branched and septate. The conidia are slimy and the
Fusarium wilt (caused by F. oxysporum), Septoria leaf spot (caused microconidia are one celled, oval or comma shaped, pyriform or
by S. lycopersici), and Phomopsis blight (caused by P. vexans). Sinha elongated, hyaline or pale, spindale shaped, and pointed (Correll et
and Singh (1983), found that sulfur fully inhibited the growth of al., 1986). The identification of A. rabiei was possible through the
Aspergillus parasiticus. Saxena et al. (1988), used 200 ppm sulfur pycnidia which were brown, thick walled and immersed in blight.
and observed a growth inhibition of 22 fungi genera. Toledo et al. Conidia are bicelled, obovoid, hyaline to palebrown. Ascochyta blight
(2009), reported an inhibited mycelial growth of A. solani under the is characterized by their spore size (Chen et al., 2004). The
silicea influence. The current study aimed to evaluate the in vitro identification of P. vexans was possible through its conidia which are
efficacy of homeopathic medicine, i.e., A. montana, T. occidentalis, hyaline, single celled, and globose to ovoid. These conidia were of
sulfur, and silicea at different concentrations (50, 100, 150, and 200 two types: alpha conidia avoid to fusoid, and beta conidia
ppm) against (a) F. oxysporum f. sp. lycopersici, (b) A. solani, (c) A. (stylospores), filiform, curved at the apex, like a walking stick, and
rabiei, (d) S. lycopersici, and (e) P. vexans. biguttulate (Dhingra and Sinclair, 1985). The identification of S.
 3

lycopersici was possible through its conidia which are hyaline, filiform montana 100 ppm; T11: A. montana 150 ppm; T12: A. montana
and multiseptated. Its pycnidia were dark, separate, globose, 200 ppm; T13: Silicea 50 ppm; T14: Silicea 100 ppm; T15: Silicea
immersed or papillate (da Costa et al., 2022). 150 ppm; T16: Silicea 200 ppm; T17: Control.
2.4 Maintenance of the cultures 2.6 Statistical analysis
The fungal pathogen was subcultured on PDA slants and allowed to Each treatment was replicated thrice and values were means ± SE.
grow at 25 ± 1oC for 10 days. Samples were stored in the refrigerator The data were computed using SPSS software version 21.
(at 4oC) and subcultured once at regular intervals of a month.
3. Results
2.5 Effect of different homeopathic medicine against major
3.1 Efficacy of homeopathic medicine against F. oxysporum f.
plant pathogenic fungi
sp. lycopersici (in vitro)
The efficacy of A. montana, T. occidentalis, sulfur, and silicea at The effect of four homeopathic medicine, i.e., A. montana, T.
different concentrations of 50, 100, 150, and 200 ppm was evaluated occidentalis, sulfur, and silicea against F. oxysporum f. sp. lycopersici
in PDA medium using poisoned food technique against different was evaluated at different concentrations (50, 100, 150, and 200
pathogens in the laboratory. The homeopathic medicine was tested ppm) under in vitro condition. Results showed a significant inhibition
with three replications per treatment. The fungus was grown on of F. oxysporum f. sp. lycopersici mycelial growth in all four tested
PDA medium for 12 days before setting up the experiment. The PDA
concentrations over control (Table 1, Figure1). Among all treatments,
medium was prepared and melted, and homeopathic medicine was the minimal radial growth of mycelium was recorded in plates treated
added to the melted medium at the required concentrations. 20 ml of with T. occidentalis at 8 days after inoculation (DAI). The highest
poisoned medium was poured into sterilized petri-plates. The mycelial growth inhibition over control was observed with T.
suitable check was maintained without the addition of homeopathic occidentalis at 200 ppm (73.73%), followed by sulfur at 200 ppm
medicine (control). A mycelial disc of 5 mm was taken from the (72.35%), and T. occidentalis at 150 ppm (70.91%), corresponding,
periphery of the old colony. The actively growing hyphal tip was respectively, to 22.80 mm, 23.00 mm, and 25.25 mm mycelial growth
removed by cork borer and placed in the center of poisoned petri- at 8 DAI. The lowest mycelial growth inhibition was attributed to
plates incubated at 25 ± 1oC until the control plate was full. The silicea at 50 and 100 ppm (29.60% and 36.12%, respectively),
radial growth of the fungus on the poisoned medium was measured corresponding, respectively, to 61.10 mm and 55.44 mm mycelial
on a daily basis up to 8 days of inoculation. The diameter of the growth at 8 DAI compared to the control. Promising inhibition
colony was measured in two directions and the average was recorded percentages of F. oxysporum f. sp. lycopersici mycelial growth were
to find out the growth of concerning pathogens. All experiments observable with A. montana at 150 and 200 ppm compared to the
were carried out in triplicate and the per cent reduction of mycelial control (61.46 and 68.66%, corresponding to 33.45 and 27.20 mm-
growth over control was calculated using the following formula of mycelial growth at 8 DAI, respectively). Hanif and Dawar (2015),
Vincent (1927): reported that F. oxysporum, R. solani and M. phaseolina was
significantly inhibited by A. montana and T. occidentalis, and the
Dc – D t growth parameters and yield of crops were improved. 200 ppm
Per cent decrease over control = ×100
Dc Arsenicum album reduced by 88% the leaf rot of betelvine disease
caused by Phytophthora spp. Hanif and Dawar (2016). Dogra (2006),
where, Dc is the average diameter of fungal growth in control and Dt observed the antifungal activity of panchgavya against major soil-
is the average diameter of fungal growth in treatment. borne fungi. He found that mycelial bits, dipped for 12 h in
Treatments were as follows: T1: Sulfur 50 ppm; T2: Sulfur 100 panchgavya, caused more than 90% inhibition of F. oxysporum f. sp.
ppm; T3: Sulfur 150 ppm; T4: Sulfur 200 ppm; T5: T. occidentalis pisi and R. solani f. sp. pisi, and 100% inhibition of S. rolfsii, S.
50 ppm; T6: T. occidentalis 100 ppm; T7: T. occidentalis 150 ppm; sclerotiorum and R. solani seeds treated with homeopathic drugs.
T8: T. occidentalis 200 ppm; T9: A. montana 50 ppm; T10: A. Besides that, plant growth parameters were improved.

Table 1: Effect of homeopathic medicine at the different concentrations on radial mycelial growth against
F. oxysporum f. sp. lycopersici
Treatments Radial mycelial growth (mm) at different days Inhibition (%) over control
2 days 4 days 6 days 8 days
T1 10.12 19.30 30.40 41.74 51.92
T2 8.90 16.70 23.27 39.10 54.95
T3 5.60 13.46 20.50 31.38 63.84
T4 3.20 7.50 13.05 23.00 72.35
T5 7.06 15.40 25.66 37.60 56.68
T6 6.03 11.06 19.62 30.87 64.43
T7 5.80 10.50 18.76 25.25 70.91
4

T8 4.30 8.92 17.40 22.80 73.73

T9 12.40 24.90 39.70 53.50 38.36
T10 10.70 20.35 30.05 51.03 41.20
T11 6.10 11.60 25.76 33.45 61.46
T12 4.75 9.15 18.20 27.20 68.66
T13 14.70 29.40 46.05 61.10 29.60
T14 13.65 25.05 40.40 55.44 36.12
T15 6.25 15.75 26.44 35.40 59.21
T16 4.80 9.55 19.90 29.70 65.78
T17 21.10 40.50 63.05 86.80 -
SE 0.368 0.714 1.142 1.612 -
CD at 1 % 1.420 2.747 4.407 6.211 -

T1: Sulfur 50 ppm; T2: Sulfur 100 ppm; T3: Sulfur 150 ppm; T4: Sulfur 200 ppm; T5: T. occidentalis 50 ppm; T6: T.
occidentalis 100 ppm; T7: T. occidentalis 150 ppm; T8: T. occidentalis 200 ppm; T9: A. montana 50 ppm; T10: A. montana
100 ppm; T11: A. montana 150 ppm; T12: A. montana 200 ppm; T13: Silicea 50 ppm; T14: Silicea 100 ppm; T15: Silicea
150 ppm; T16: Silicea 200 ppm; T17: Control.

Thuja @ 50 ppm Sulphur @ 50 ppm Arnica @ 50 ppm Silicea @ 50 ppm

Thuja @ 100 ppm Sulphur @ 100 ppm Arnica @ 50 ppm Silicea @ 100 ppm

Thuja @ 150 ppm Sulphur @ 150 ppm Arnica @ 50 ppm Silicea @ 150 ppm
 5

Thuja @ 200 ppm Sulphur @ 200 ppm Arnica @ 50 ppm Silicea @ 200 ppm

Co ntr o l

Figure 1: Effect of homeopathic medicine at different concentrations on radial mycelial growth of Fusarium wil t of tomato
caused by F. oxysporum f. sp. lycopersici
3.2 Efficacy of homeopathic medicine against A. solani (in vitro) to silicea at 50 ppm (39.42%), corresponding to a 47.13 mm mycelial
growth at 8 DAI.
The effect of four homeopathic medicine, i.e., T. occidentalis, sulfur,
A. montana, and silicea against A. solani was evaluated at different Dahiwale and Suryawanshi (2010), reported the antifungal activities
concentrations (50, 100, 150, and 200 ppm) under in vitro condition. of homeopathic medicines, i.e., T. occidentalis, sulfur, A. montana,
Results showed a significant inhibition of the mycelial growth of A. A. phosphoricum, Spongia tosta, and Chelidonium majus. These
solani in all four tested concentrations over control (Table 2, Figure treatments were tested individually and in mixture with mancozeb
2). Among all treatments, the minimal radial growth of the mycelium against the mycelial growth of A. alternata using potato dextrose
was recorded in plates treated with T. occidentalis at 8 DAI. The agar (PDA) medium via food poisoning method. Dua and Atri (2004),
highest mycelial growth inhibition over control was observed with reported the antifungal effect of homeopathic medicine, i.e., T.
T. occidentalis at 200 ppm (76.73%), followed by A. montana at 200 occidentalis and Lycopodium spp. against A. solani, the main reason
ppm (75.35%), sulfur at 200 ppm (74.29%), and T. occidentalis at of tomato and potato early blight disease. Patil and Suryawanshi
150 ppm (74.03%), corresponding, respectively, to 18.10 mm, 19.17 (2014), noticed the potential control of strawberry fruit rot caused
mm, 20.00 mm, and 20.20 mm mycelial growth at 8 DAI. The lowest by A. alternata using different homeopathic medicines with promising
mycelial growth inhibition compared to the control was attributed effects on vegetative and reproductive growth.
Table 2: Effect of homeopathic medicine at different concentrations on radial mycelial growth against early
blight disease of tomato caused by A. solani
Treatments Radial mycelial growth (mm) at different days Inhibition (%) over control
2 days 4 days 6 days 8 days
T1 12.40 17.24 29.06 36.00 53.72
T2 10.44 13.90 22.30 29.30 62.33
T3 8.93 12.29 18.95 22.50 71.07
T4 6.06 10.22 15.63 20.00 74.29
T5 8.12 12.10 19.10 26.66 65.73
T6 7.26 11.06 18.78 25.63 67.05
T7 6.93 10.10 16.77 20.20 74.03
T8 4.70 8.90 13.50 18.10 76.73
T9 11.29 16.78 26.42 32.44 58.30
6

T10 9.70 12.92 20.97 27.90 64.13

T11 7.72 11,23 17.90 22.00 71.12

T12 5.95 9.82 14.44 19.17 75.35

T13 16.30 28.66 38.72 47.13 39.42

T14 14.50 18.75 36.07 37.50 51.79

T15 9.95 11.77 26.33 24.40 69.92

T16 6.90 11.41 17.08 21.47 72.40

T17 24.50 39.70 56.50 77.80 -

SE 0.411 0.642 0.961 1.206 -

CD at 1 % 1.587 2.653 4.475 6.180 -

T1: Sulfur 50 ppm; T2: Sulfur 100 ppm; T3: Sulfur 150 ppm; T4: Sulfur 200 ppm; T5: T. occidentalis 50 ppm; T6: T.
occidentalis 100 ppm; T7: T. occidentalis 150 ppm; T8: T. occidentalis 200 ppm; T9: A. montana 50 ppm; T10: A.
montana 100 ppm; T11: A. montana 150 ppm; T12: A. montana 200 ppm; T13: Silicea 50 ppm; T14: Silicea 100 ppm;
T15: Silicea 150 ppm; T16: Silicea 200 ppm; T17: Control.

Thuja @ 50 ppm Sulphur @ 50 ppm Arnica @ 50 ppm Silicea @ 50 ppm

Thuja @ 100 ppm Sulphur @ 100 ppm Arnica @ 50 ppm Silicea @ 100 ppm

Thuja @ 150 ppm Sulphur @ 150 ppm Arnica @ 50 ppm Silicea @ 150 ppm
 7

Thuja @ 200 ppm Sulphur @ 200 ppm Arnica @ 50 ppm Silicea @ 200 ppm

Control
Figure 2: Effect of homeopathic medicine at different concentrations on radial mycelial growth of early blight disease of tomato
caused by A. solani

3.3 Efficacy of homeopathic medicine against A. rabiei (in vitro) lowest mycelial growth inhibition over control was attributed to
silicea at 50 ppm (34.99%), corresponding to a 57.73 mm mycelial
The effect of four homeopathic medicine, i.e., T. occidentalis, sulfur,
growth at 8 DAI.
A. montana, and silicea was evaluated against A. rabiei at different
concentrations (50, 100, 150, and 200 ppm) under in vitro condition. Ganzera et al. (2008) and Sugier et al. (2009), reported that Arnica
Results showed a significant inhibition of the mycelial growth of A. spp. contain essential oils, terpenoids, flavonoids, and phenolic acids,
rabiei in all four tested concentrations over control (Table 3, Figure especially pseudoguaianolide sesquiterpenes (0.2-0.8%) initially
3). Among all treatments, the minimal radial growth of mycelium located in the flower head of A. montana, which exhibits antioxidant
was recorded in plates treated with A. montana at 8 DAI. The highest and antifungal activities against plant pathogens. Arie (2019), reported
mycelial growth inhibition over control was observed with A. montana the effect of four homeopathic drugs, i.e., Thuja spp., Calcarea
at 200 ppm (64.22%) followed by T. occidentalis at 200 ppm carbonica, nitric acid and sulfur against A. alternata,
(63.77%), A. montana at 150 ppm (62.18%), and T. occidentalis at Helminthosporium ramera, and F. oxysparum. The author outlined
150 ppm (61.33%), corresponding, respectively, to 31.66 mm, 32.06 that Thuja spp., nitric acid and sulfur completely inhibited the growth
mm, 33.47 mm, and 34.22 mm mycelial growth at 8 DAI. The of all fungi.
Table 3: Effect of homeopathic medicine at different concentrations on radial mycelial growth against Ascochyta
blight of chickpea caused by A. rabiei
Treatments Radial mycelial growth (mm) at different days Inhibition (%) over control
2 days 4 days 6 days 8 days

T1 13.70 26.23 41.16 57.73 34.99

T2 12.08 21.43 33.43 49.58 43.97

T3 11.53 20.66 32.62 47.52 46.30

T4 9.62 16.57 27.19 38.47 56.53

T5 10.10 18.17 28.06 43.41 50.94

T6 9.88 17.70 26.50 39.57 55.28

T7 8.55 15.42 25.06 34.22 61.33

8

T8 7.40 13.87 23.80 32.06 63.77

T9 9.98 17.78 27.87 40.38 54.37
T10 8.05 15.02 25.77 35.50 59.88
T11 7.84 14.82 24.50 33.47 62.18
T12 6.08 12.16 22.20 31.66 64.22
T13 12.50 24.67 38.87 50.60 42.82
T14 11.20 19.60 31.44 46.16 47.84
T15 10.46 16.05 28.32 44.54 49.67
T16 8.15 15.04 25.76 36.36 58.91
T17 32.45 48.64 69.10 88.50 -
SE 0.436 0.751 1.168 1.605 -
CD at 1 % 1.738 2.629 4.572 6.134 -
T1: Sulfur 50 ppm; T2: Sulfur 100 ppm; T3: Sulfur 150 ppm; T4: Sulfur 200 ppm; T5: T. occidentalis 50 ppm; T6: T.
occidentalis 100 ppm; T7: T. occidentalis 150 ppm; T8: T. occidentalis 200 ppm; T9: A. montana 50 ppm; T10: A. montana
100 ppm; T11: A. montana 150 ppm; T12: A. montana 200 ppm; T13: Silicea 50 ppm; T14: Silicea 100 ppm; T15: Silicea
150 ppm; T16: Silicea 200 ppm; T17: Control.

Thuja @ 50 ppm Sulphur @ 50 ppm Arnica @ 50 ppm Silicea @ 50 ppm

Thuja @ 100 ppm Sulphur @ 100 ppm Arnica @ 50 ppm Silicea @ 100 ppm

Thuja @ 150 ppm Sulphur @ 150 ppm Arnica @ 50 ppm Silicea @ 150 ppm
 9

Thuja @ 200 ppm Sulphur @ 200 ppm Arnica @ 50 ppm Silicea @ 200 ppm

Co ntr o l

Figure 3: Effect of homeopathic medicine at different concentrations on radial mycelial growth of Ascochyta bl ight of chickpea
caused by A. rabiei.

3.4 Efficacy of homeopathic medicine against S. lycopersici (in and 37.11 mm mycelial growth at 8 DAI. The lowest mycelial growth
vitro) inhibition over control was attributed to A. montana at 50 ppm
(29.56%), corresponding to a 61.12 mm mycelial growth at 8 DAI.
The effect of four homeopathic medicine, i.e., T. occidentalis, sulfur,
A. mintana, and silicea was evaluated against S. lycopersici at different Hanif and Dawar (2015), evaluated the in vitro effect of homeopathic
concentrations (50, 100, 150, and 200 ppm) under in vitro condition. medicine, namely; A. montana and T. occidentalis against root rot
Results showed a significant inhibition of the mycelial growth of S. fungi (F. oxysporum, R. solani, and M. phaseolina) and outlined
lycopersici in all four tested concentrations over control (Table 4, promising values. Khanna and Chandra (1989), reported a positive
effect of homeopathic medicines, i.e., Lycopodium spp., Thuja spp.,
Figure 4). Among all treatments, the minimal radial growth of Arsenicum spp., and Zincum spp. against F. moniliforme, A. alternata,
mycelium was recorded in plates treated with silicea at 8 DAI. The Gloeosporium psidii, Colletotrichum gloeosporioides, and Pestalotia
highest mycelial growth inhibition over control was observed with spp. succeeding in the control of these fruit rot pathogens. Few
silicea at 200 ppm (59.77%), followed by sulfur at 200 ppm (58.95%), researchers have reported the same aspects in controlling plant
silicea at 150 ppm (57.50%), and sulfur at 150 ppm (57.23%), pathogens. For instance, Saxena et al. (1988) reported the antifungal
corresponding, respectively, to 34.91 mm, 35.62 mm, 36.88 mm, activity of T. occidentalis used for okra seed treatment.

Table 4:Effect of homeopathic medicine at different concentrations on radial mycelial growth against Septoria
leaf spot of tomato caused by S. lycopersici
Treatments Radial mycelial growth (mm) at different days Inhibition (%) over control
2 days 4 days 6 days 8 days

T1 11.50 22.90 32.37 43.60 49.75

T2 10.10 18.57 23.45 39.44 54.55

T3 8.10 14.08 22.60 37.11 57.23
T4 7.25 12.27 21.93 35.62 58.95

T5 13.67 24.47 36.47 45.61 47.44

T6 12.44 23.42 34.76 44.72 48.46

T7 10.12 19.15 29.18 42.21 51.35

T8 9.41 17.77 26.87 40.32 53.53

T9 15.43 27.67 39.40 61.12 29.56

10

T10 13.12 25.70 36.31 45.63 47.41

T11 11.29 23.68 34.47 43.11 50.32

T12 10.53 13.10 28.77 41.22 52.50

T13 9.93 18.50 28.42 41.81 51.82

T14 8.15 16.60 22.58 38.40 55.75

T15 7.87 13.88 21.10 36.88 57.50

T16 6.72 12.66 20.23 34.91 59.77

T17 22.17 39.56 61.87 86.78 -

SE 0.407 0.750 1.124 1.632 -

CD at 1 % 1.634 2.256 4.214 6.381 -

T1: Sulfur 50 ppm; T2: Sulfur 100 ppm; T3: Sulfur 150 ppm; T4: Sulfur 200 ppm; T5: T. occidentalis 50 ppm; T6: T.
occidentalis 100 ppm; T7: T. occidentalis 150 ppm; T8: T. occidentalis 200 ppm; T9: A. montana 50 ppm; T10: A. montana
100 ppm; T11: A. montana 150 ppm; T12: A. montana 200 ppm; T13: Silicea 50 ppm; T14: Silicea 100 ppm; T15: Silicea
150 ppm; T16: Silicea 200 ppm; T17: Control.

Thuja @ 50 ppm Sulphur @ 50 ppm Arnica @ 50 ppm Silicea @ 50 ppm

Thuja @ 100 ppm Sulphur @ 100 ppm Arnica @ 50 ppm Silicea @ 100 ppm

Thuja @ 150 ppm Sulphur @ 150 ppm Arnica @ 50 ppm Silicea @ 150 ppm
 11

Thuja @ 200 ppm Sulphur @ 200 ppm Arnica @ 50 ppm Silicea @ 200 ppm

Co ntr o l

Figure 4: Effect of homeopathic medicine at different concentrations on radial mycelial growth of Septoria leaf spot of tomato
caused by S. lycopersici.

3.5 Efficacy of homeopathic medicine against P. vexans (in vitro) and Sulfur at 150 ppm (50.93%), corresponding respectively to
38.12 mm, 39.90 mm, and 40.50 mm mycelial growth at 8 DAI. The
The effect of four homeopathic medicine, i.e., T. occidentalis, sulfur,
lowest mycelial growth inhibition over control was attributed to
A. montana, and silicea was evaluated against P. vexans at different
silicea at 50 ppm (38.82%), corresponding to a 50.50 mm mycelial
concentrations (50, 100, 150, and 200 ppm) under in vitro condition.
growth at 8 DAI. Asha et al. (2014), reported the effective role of
Results showed a significant inhibition of the mycelial growth of P.
T. occidentalis against A. solani, F. oxysporum sp. lycopersici, and P.
vexans in all four tested concentrations over control (Table 5, Figure
vexans fungal genera. Panda et al. (2013), treated cereal seeds with
5). Among all treatments, the minimal radial growth of mycelium
0.1% T. occidentalis. They outlined an improvement in plant growth
was recorded in plates treated with sulfur at 8 DAI. The highest
parameters, i.e., shoot length and weight, and root length and weight
mycelial growth inhibition over control was observed with sulfur at
of cereal crops. Particularly, the growth of millet and wheat root
200 ppm (53.82%), followed by T. occidentalis at 200 ppm (51.66%),
infecting fungi.
Table 5: Effect of homeopathic medicine at different concentrations on radial mycelial growth against
Phomopsis blight of brinjal caused by P. vexans
Treatments Radial mycelial growth (mm) at different days Inhibition (%) over control
2 days 4 days 6 days 8 days

T1 13.77 22.50 33.20 46.77 43.34

T2 10.20 20.70 31.09 41.87 49.27

T3 8.95 19.06 30.96 40.50 50.93

T4 7.42 18.40 29.05 38.12 53.82

T5 14.60 26.78 38.88 47.50 42.45

T6 11.05 24.70 36.50 43.12 47.76

T7 9.15 22.15 34.10 41.50 49.72

T8 7.70 21.83 32.70 39.90 51.66

T9 16.90 28.50 40.50 48.20 41.61

T10 15.85 26.80 38.78 46.50 43.67

12

T11 11.90 24.95 36.30 45.44 44.95

T12 8.50 23.10 35.12 43.50 47.30

T13 17.30 30.06 41.34 50.50 38.82

T14 16.70 27.40 39.12 47.70 42.21

T15 12.40 25.12 37.10 46.21 44.22

T16 9.10 24.50 36.35 44.72 45.82

T17 23.20 41.50 64.30 82.55 -

SE 0.485 0.913 1.358 1.686 -

CD at 1 % 1.899 2.795 4.691 6.867 -

T1: Sulfur 50 ppm; T2: Sulfur 100 ppm; T3: Sulfur 150 ppm; T4: Sulfur 200 ppm; T5: T. occidentalis 50 ppm; T6: T.
occidentalis 100 ppm; T7: T. occidentalis 150 ppm; T8: T. occidentalis 200 ppm; T9: A. montana 50 ppm; T10: A.
montana 100 ppm; T11: A. montana 150 ppm; T12: A. montana 200 ppm; T13: Silicea 50 ppm; T14: Silicea 100 ppm;
T15: Silicea 150 ppm; T16: Silicea 200 ppm; T17: Control.

Thuja @ 50 ppm Sulphur @ 50 ppm Arnica @ 50 ppm Silicea @ 50 ppm

Thuja @ 100 ppm Sulphur @ 100 ppm Arnica @ 50 ppm Silicea @ 100 ppm

Thuja @ 150 ppm Sulphur @ 150 ppm Arnica @ 50 ppm Silicea @ 150 ppm
 13

Thuja @ 200 ppm Sulphur @ 200 ppm Arnica @ 50 ppm Silicea @ 200 ppm

Co ntr o l

Figure 5: Effect of homeopathic medicine at different concentrations on radial mycelial growth of Phomopsis bl ight of brinjal
caused by P. vexans.

4. Discussion significantly inhibited mycelial growth of S. lycopersici and P. vexans,

in all four tested concentrations over control (Table 4, Figure 4 and
Effect of four homeopathic medicines, viz., T. occidentalis, sulphur, Table 5, Figure 5). The maximum inhibition of radial growth of S.
A. mintana, and silicea, at different concentrations (50, 100, 150 and lycopersici was recorded in Figure 4 treated with silicea @ 50, 100,
200 ppm) were evaluated in vitro condition and found that 150 and 200 ppm, representing as 41.81 mm, 38.80 mm, 36.88 mm
significantly inhibited mycelial growth of F. oxysporum f. sp. and 34.91 mm on 8 days after inoculation (DAI), which was inhibited
lycopersici, in all four tested concentrations over control (Table 1 59.77% mycelial growth over control as well as in Phomopsis vexans,
and Figure1). Among all these treatments, the minimum radial growth the minimum radial growth of mycelium was recorded in Figure 5
of mycelium was recorded in plates treated with 50, 100, 150 and treated with 50, 100, 150 and 200 ppm of sulphur, representing as
200 ppm of T. occidentalis, representing as 37.60 mm, 30.87 mm, 46.77 mm, 41.87 mm, 40.50 mm and 38.12 mm, at 8 days after
25.25 mm and 22.80 mm, at 8 days after inoculation (DAI), which inoculation (DAI), which was highest (53.82%) inhibition of mycelial
was highest (73.73%) inhibition of mycelial growth over control. growth over control.
Similarly, in A. solani (Table 2 and Figure 2), the maximum inhibition
of radial growth of mycelium was observed in plates treated with T. 5. Conclusion
occidentalis @ 50, 100, 150 and 200 ppm, representing as 26.66
Plants produce secondary metabolites throughout the homoeopathic
mm, 25.63 mm, 20.20 mm and 18.10 mm,on 8 days after inoculation
drug making process. These metabolites are non-toxic and non-residue
(DAI), which was highest inhibited 76.73% of mycelial growth over
forming, making them safe to the environment. This study
control.
investigated the synergistic effect of homoeopathic medications and
In A. rabiei, effect of four homeopathic medicine (T. occidentalis, microbial antagonist in preventing several fungal diseases, i.e.,
sulphur, A. mintana, and silicea) at different concentration (50, 100, Fusarium wilt, early blight of potato and Septoria leaf spot of tomato,
150 and 200 ppm) were evaluated in vitro condition and found that Ascochyta blight of chickpea, and Phomopsis blight of brinjal.
significantly inhibited mycelial growth of all four tested
Pathogens (Fusarium, Phomopsis, Septoria, Alternaria, and
concentrations over control (Table 3 and Figure 3). Among all these
Ascochyta), responsible for plant diseases were treated using
treatments, the minimum radial growth of mycelium was recorded in
homoeopathic remedies including A. montana, T. occidentalis, sulfur,
plates treated with 50, 100, 150 and 200 ppm of A. montana,
and silicea. The increased concentrations of homeopathic medicine
representing as 40.38 mm, 35.50 mm, 33.47 mm and 31.66 mm, at 8 were found to significantly reduce the radial mycelial growth of F.
days after inoculation (DAI), which was highest (64.22%) inhibition oxysporum f. sp. lycopersici, A. solani, S. lycopersici, A. rabiei, and
of mycelial growth over control. P. vexans under in vitro condition. At 8 DAI, T. occidentalis at 200
Effect of four homeopathic medicine, viz., T. occidentalis, sulphur, ppm (73.73%), T. occidentalis at 200 ppm (76.73%), A. montana at
A. mintana, and silicea, at different concentration (50, 100, 150 and 200 ppm (64.22%), silicea at 200 ppm (59.77%), and sulfur at 200
200 ppm) were evaluated in vitro condition and found that ppm (53.82%) showed the maximum mycelial growth inhibition
14

compared to the control. The minimal radial growth for these five Conforti, A.; Bertani, S.; Metelmann, H.; Chirumbolo, S.; Lussignoli, S. and
fungal species was 22.80 mm, 18.10 mm, 31.66 mm, 34.91 mm, and Bellavite, P. (1997). Experimental studies of the anti-inflammatory
38.12 mm, respectively. These homeopathic remedies can, therefore, activity of a homeopathic preparation. Biol. Ther., 15 :28-31.
help plants defend themselves against fungus and other pathogen Correll, J. C.; Puhalla, J. E. and Schneider, R. W. (1986). Identification of
attack. Further, research should investigate the efficacy of used Fusarium oxysporum. f. sp. apii on the basis of colony size,
homeopathic remedies against other fungal species affecting virulence, and vegetative compatibility. Phytopathology, 76 (4):
horticultural crops. 396-400.

Conflict of interest da Costa, C. A.; Lourenço Jr, V.; Santiago, M. F.; Veloso, J. S. and Reis, A. (2022).
Molecular phylogenetic, morphological, and pathogenic analyses
The authors declare no conflicts of interest relevant to this article. reveal a single clonal population of Septoria lycopersici with a
narrower host range in Brazil. Plant Pathology, 71 (3):621-633.
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