Pharmaceutical Biology

ISSN: 1388-0209 (Print) 1744-5116 (Online) Journal homepage: https://www.tandfonline.com/loi/iphb20

Antifungal Activity of Some Plant Extracts Against

Fungal Pathogens of Tea (Camellia sinensis.)

D. Saha, S. Dasgupta & A. Saha

To cite this article: D. Saha, S. Dasgupta & A. Saha (2005) Antifungal Activity of Some Plant
Extracts Against Fungal Pathogens of Tea (Camellia�sinensis.), Pharmaceutical Biology, 43:1,
87-91, DOI: 10.1080/13880200590903426

To link to this article: https://doi.org/10.1080/13880200590903426

Published online: 07 Oct 2008.

Submit your article to this journal

Article views: 12820

View related articles

Citing articles: 9 View citing articles

Full Terms & Conditions of access and use can be found at

https://www.tandfonline.com/action/journalInformation?journalCode=iphb20
Pharmaceutical Biology
2005, Vol. 43, No. 1, pp. 87–91
Antifungal Activity of Some Plant Extracts Against Fungal
Pathogens of Tea (Camellia sinensis)
D. Saha, S. Dasgupta, and A. Saha
Mycology and Plant Pathology Laboratory, Department of Botany, University of North Bengal, Siliguri, India
Abstract
In an approach toward the development of ecofriendly
antifungal compounds for controlling major foliar fungal
diseases of tea, ethanol and aqueous extracts of 30 plants
belonging to 20 different families collected from sub-
Himalayan West Bengal (India) were tested against
the pathogens Pestalotiopsis theae (Saw.) Stey., Colle-
totrichum camelliae Mess., Curvularia eragrostidis
(P. Hennings) Meyer, and Botryodiplodia theobromae
Patouiilard. Spore germination technique was followed
for evaluation of antifungal properties. Results showed
that ethanol and aqueous extracts of Allium sativum L.,
Datura metel L., Dryopteris filix-mas (L.) Schott,
Zingiber officinale Rosc., Smilax zeylanica L., Azadir-
achta indica, A. Joss. and Curcuma longa L. recorded
100% inhibition of spore germination. The antifungal
component from these plants may be used in developing
novel fungicides for tea gardens.
Keywords: Antifungal activity, Camellia sinensis, plant
extracts.
Introduction
Tea is a major plantation crop of India and is cultivated
extensively throughout north east India. Fungal patho-
gens present a significant threat to tea leaves that
requires use of chemical fungicides. However, the use
of chemical fungicides for tea plants has increasingly
become unpopular due to an all-round awareness of its
polluting effects, leading to tighter health and environ-
mental regulations. This in turn has created a need for
alternative sourcing of fungicidal agents that can be
developed for treatment of fungal tea-leaf diseases.
Accepted: October 4, 2004
Address correspondence to: D. Saha, Mycology and Plant Pathology Laboratory, Department of Botany, University of North Bengal,
Siliguri 734430, India. E-mail: drishtanta@yahoo.com
DOI: 10.1080/13880200590903426
The traditional practice of using plant preparations to
combat fungal infections has gained attention, and
currently the focus is on detection of new antifungal
components from plants that have no negative effect on
the environment or on animal and human systems. Several
authors have reported antifungal activity of plant extracts
against pathogens of rice, tomato, wheat, pea, and other
important crops (Rana et al., 1999; Sindhan et al., 1999;
Hu et al., 2001). However, such reports involving tea
pathogens are very few (Chakraborty et al., 1991).
This paper presents a study on the in vitro effect of
aqueous and ethanol extracts of 30 plant species against
four major foliar fungal pathogens of tea. This is an
approach to identify antifungal elements in natural
sources that may be used for controlling tea-leaf diseases.
Materials and Methods
Plant material
The extracts of 30 species of plants (29 angiosperms and
one pteridophyte) belonging to 20 different families have
been evaluated for their antifungal activity. Table 1
shows a list of the plants along with their local names
and traditional uses (Rastogi & Mehrotra, 1995;
Chatterjee & Pakrashi, 1997; Chopra et al., 1996). Some
of the plant materials were collected from forest areas of
the Mahananda Wild Life Sanctuary located in the terai
region of the eastern Himalayas. Others were collected
from local areas within and outside the campus of the
University of North Bengal. Voucher specimens have
been deposited in the departmental herbarium of the
Department of Botany, University of North Bengal.
# 2005 Taylor & Francis Ltd.
 Table 1. List of plants tested in this study.

88
Plant species tested Local name Family Plant parts used Traditional uses

Allium cepa L. Piyaj Alliaceae Bulb Stimulant, diuretic, expectorant, in flatulence and dysentery
Allium sativum L. Lasan Alliaceae Bulb Juice used as rubefacient in skin diseases, used in
atonic dyspepsia, flatulence, and colic
Andrographis peniculata Nees Kalmegh Acanthaceae Leaf Useful as stomachic, febrifuge, antihelmintic, in
dysentery and dyspepsia
Artocarpus heterophylus Lam. Kathal Moraceae Leaf Used in skin diseases, antidote to snake bite
Azadirachta indica A. Juss. Neem Meliaceae Leaf Applied to boils as poultice, decoction antiseptic,
used in ulcers and eczema
Bougainvillea spectabilis Willd. Bogenvila Nyctaginaceae Leaf Pinitol from leaves produce hypoglycemia in mice
Cassia tora L. Chakunda Caesalpinoideae Aerial parts Cure ringworm, itches, eczema, and other skin diseases
Catharanthus roseus (L.) G. Don Nayantara Apocynaceae Leaf Infusion given in menorrhagia
Curcuma longa L. Haldi Zingiberaceae Rhizome Stimulant, blood purifier, externally applied to sprains
and wounds, juice is antihelmintic, used as antiparasitic
for many skin infections
Datura metel L. Dhutura Solanaceae Leaf Antiseptic, anodyne, antispasmodic
Dryopteris filix-mas (L.) Schott Dhekisag Polypodiaceae Aerial parts Fern extract taeniacidal against dwarf tapeworm
Emblica officianilis Gaertn. Amla Euphorbiaceae Leaf Juice applied externally to ulcers, infusion useful in
chronic dysentery
Euphorbia hirta L. Boro-keruie Euphorbiaceae Aerial parts Juice is given in colic dysentery, decoction in asthma
and bronchial infection
Gmellina philippensis Cham — Verbenaceae Leaf Ethanol extract spasmolytic, diuretic, and anticancer
Hyptis suaveolens (L.) Poit. Bilati Tulsi Labiatae Leaf Juice given in colic
Jatropha curcas L. Bagbherenda Euphorbiaceae Leaf Oil is applied in chronic rheumatism and skin diseases
Lantana camara L. var. aculeate Moldenke Guye genda Verbenaceae Leaf Antiseptic, antispasmodic, carminative, promotes healing
of wounds
Leea indica (Burm.f.) Merr. Kurkurjiwah Leeaceae Leaf Ointment prepared from roasted leaves relieves vertigo,
juice is digestive
Leucus indica (L.) R. Br. ex Vatke Halkasa Labiatae Aerial parts Paste of fresh leaves applied externally to dermatosis
and old sores
Moringa oleifera Lamk. Sajna Moringaceae Leaf Used as purgative and for cough
Murraya koenigi (L.) Spreng. Barsunga Rutaceae Leaf Bruised and applied locally to eruptions and poisonous bite
Nerium indicum Mill. Karabi Apocynaceae Leaf Paste of leaves used in ringworm and other skin diseases,
juice applied in syphilitic ulcers
Ocimum sanctum L. Tulsi Labiatae Leaf Fresh leaves bruised and applied externally for curing
ring worm and other skin diseases
Piper peepuloides Roxb. — Piperaceae Leaf Juice is diaphoretic, expectorant, and stimulant
Plumeria rubra L. forma acuminate (Ait.) Gorur champa Apocynaceae Leaf Febrifuge, paste applied as a poultice
Sant. & Irani ex Shah
Psidium guajava L. Peyara Myrteceae Leaf Used as a gargle for mouthwash in swollen gum,
locally applied to ulcers
Ricinus comunis L. Rehri Euphorbiaceae Leaf Applied externally to boils and sores
Smilax zeylanica L. Kumarika Smilacaceae Leaf Used in rheumatism and pain, used in bloodless dysentery
Tagetes erecta L. Genda Compositae Aerial parts Externally applied to boils and carbuncles, anodyne
Zingiber officinale Rosc. Ada Zingiberaceae Rhizome Used in dyspepsia and flatulent colic
 Antifungal activity of plant extracts against pathogens of tea 89

Table 2. Effect of different plant extracts on spore germination of Pestalotiopsis theae (P.t.), Colletotrichum camelliae (C.c.),
Curvularia eragrostridis (C. e.), and Botryodiplodia theobrome (B. t.).

% Inhibition

Aqueous extract Ethanol extract

Plant species tested P. t. C.c. C. e. B. t. P. t. C.c. C. e. B. t.

Allium cepa L. 26.4 67.5 42.7 96.8 41.5 17.6 72.1 61.9
Allium sativum L. 100.0 100.0 96.8 85.5 100.0 100.0 88.5 80.2
Andrographis peniculata Nees 23.3 47.4 36.5 60.2 32.2 44.0 16.7 38.4
Artocarpus heterophylus Lam. 12.8 31.3 31.5 36.8 25.5 4.6 14.7 54.5
Azadirachta indica A. Juss. 100 39.8 63.0 39.2 44.7 76.0 74.4 95.2
Bougainvillea spectabilis Willd. 60.4 42.1 69.7 90.3 62.4 54.0 54.7 64.2
Cassia tora L. 64.6 35.6 23.5 38.5 76.0 40.3 63.5 56.4
Catharanthus roseus (L.) G. Don 38.7 18.7 87.5 61.2 18.5 42.4 40.2 45.4
Curcuma longa L. 62.7 99.7 98.2 96.7 58.8 100 85.5 52.2
Datura metel L. 100 88.4 85.4 71.4 100 100 92.2 79.2
Dryopteris filix-mas (L.) Schott 98.6 100 100 93.8 100 99.4 96.1 83.2
Emblica officianilis Gaertn. 28.2 38.4 69.4 66.7 30.2 47.0 87.7 65.3
Euphorbia hirta L. 10.8 12.5 10.2 7.2 16.4 12.6 7.4 12.5
Gmellina philippensis Cham 42.4 26.4 35.0 73.6 36.4 31.1 64.8 43.1
Hyptis suaveolens (L.) Poit. 11.5 21.6 40.2 9.7 28.6 15.8 12.7 57.1
Jatropha curcas L. 78.4 48.7 12.5 34.2 19.7 27.6 14.8 46.5
Lantana camara L. var. aculeate Moldenke 36.6 19 62.1 15.4 39.4 44.8 53.1 71.4
Leea indica (Burm.f.) Merr. 76.8 22.4 12.5 16.8 91.7 26.6 97.6 74.7
Leucus indica (L.) R. Br. ex Vatke 7.14 15.3 62.5 14.4 14.1 9.4 62.2 66.7
Moringa oleifera Lamk. 67 11.1 9.4 5.2 56.4 7.3 14.4 20.7
Murraya koenigi (L.) Spreng. 12.0 52.3 54.7 88.4 43.7 36.2 23.5 44.6
Nerium indicum Mill. 51.42 44.5 34.2 28.5 65 29.4 79.2 65.3
Ocimum sanctum L. 32.8 20.3 24.6 19.5 16.8 6.4 33.4 28.4
Piper peepuloides Roxb. 44.3 46.5 58.1 81.4 94.5 67.4 36.2 32.1
Plumeria rubra L. forma acuminate 54 25.7 54.2 45.4 24.7 22 32.4 53.7
(Ait.) Sant. & Irani ex Shah
Psidium guajava L. 96.7 75.6 36.8 42.5 99 79 88.4 73.7
Ricinus comunis L. 18.5 31 36.2 25.4 12.7 42.8 22.5 14.6
Smilax zeylanica L. 79.6 51.4 63.5 42.3 100 45.6 55.8 74.5
Tagetes erecta L. 9.67 8.4 57.2 21.4 37.4 11.5 8.4 11.4
Zingiber officinale Rosc. 100 88.9 63.6 68.1 100 96 81.0 72.8
SED 17.8 12 15.4 12.4 16.8 18.2 18.4 15.2

Preparation of extracts tiopsis theae (Saw.) Stey., Colletotrichum camelliae Mass.

[imperfect state of Glomerella cingulata (Stonem) S. & v.
Fresh plant materials were washed thoroughly with ster-
S.], Curvularia eragrostidis (P. Hennings) Meyer, and
ile distilled water and allowed to dry at room tempera-
Botryodiplodia theobromae Patouiilard were isolated
ture. The dried materials were ground and extracted
from infected leaves of tea plants. They were identified
separately with sterile distilled water and ethanol
and, after verification of Koch’s postulates, are
(0.5 g=ml). The extracts were filtered through double-
maintained in the laboratory in potato destrose agar
layered cheesecloth and centrifuged at 10,000  g for
(PDA) slants at 4C.
30 min. The supernatants of aqueous extracts were steri-
lized by passing through a Millipore filter (0.2 mm). All
extracts were stored at 4C and used for bioassay. Effect of plant extracts on spore germination
Plant extracts (30 ml) were placed at two spots 3-cm apart
on each microscopic slide taken in duplicate. Spore sus-
Fungal culture
pension was prepared from 12-day-old cultures of the
The plant extracts were tested against four important fungus in sterile distilled water and added (30 ml) to the
fungal pathogens. The four fungal pathogens, Pestalo- plant extracts. Control slides contained appropriate
90 D. Saha et al.
solvents in place of plant extracts. The slides were incu-
bated at 30C in a humid chamber. After 24 h, they were
stained with cotton blue=lactophenol mixture and
observed (500 spores) under the microscope. Germi-
nation, if any, was noted, and the percentage inhibition
was calculated.
Statistical analysis
The mean values of the data for inhibition of spore ger-
mination of each extract were compared with control
using Student’s t-test.
Results
Table 2 shows the effect of ethanol and aqueous extracts
of 30 plants on spore germination of four major foliar
fungal pathogens of tea. Among these, extracts from nine
plants showed significant inhibition against all the four
tested fungi. Spore germination of Pestalotiopsis theae
was totally inhibited (100% inhibition) by ethanol and
aqueous extracts from Datura metel, Allium sativum,
and Zingiber officinale. In addition, aqueous leaf extracts
from Azadirnachta indica and ethanol extracts from
Smilax zeylanica and Dryopteris filix-mas were equally
effective. The germination of spores of Colletotrichum
camelliae was totally inhibited by ethanol and aqueous
extracts from Allium sativum bulbs and also by aqueous
extracts from Dryopteris filix-mas and ethanol extracts
from Curcuma longa and Datura metel plants. Curcuma
longa extracts were similarly effective against Curvularia
eragrostidis. Spore germination of Botryodiplodia
theobromae was completely inhibited only by ethanol
extracts from Allium sativum bulbs. However, extracts
from several plants, which were effective against other
fungal pathogens, recorded more than 90% inhibition
of spore germination. Altogether, barring a few excep-
tions, the ethanol extracts seemed to be more effective
than the aqueous extracts.
Discussion
Considering the need for alternative biorational fungi-
cides in tea gardens, it was believed to be worthwhile
to evaluate the antifungal effects of locally available
plant extracts. The results were encouraging, as several
plants showed total inhibition of spore germination.
Because, for the pathogenic fungi, spore germination is
a determining factor at the onset of host colonization,
many of these extracts have a definite potential for new
effective fungicides.
Among the different plants whose extracts were
found to be effective, Allium sativum bulbs showed
maximum potential because both its aqueous and
ethanol extracts showed either 100% or more than 90%
inhibition of spore germination of all the fungal patho-
gens. Antifungal properties of Allium sativum are well-
known (Muhsin et al., 2000; Srinivasan et al., 2001),
especially against human pathogens (Samuel et al.,
2000) and also against plant pathogens (Sindhan et al.,
1999). It contains different antimicrobial components
like allicin, E-and Z-ajoene, iso-E-10-devinylajoene,
and so forth, which are effective against bacteria, yeasts,
and phytopathogenic fungi (Prithiviraj et al., 1998;
Yoshida et al., 1999).
Leaf extracts from Datura metel also showed high
antifungal efficiency against all the tested pathogens. In
a field study, Asha and Kannabiran (2001) observed that
aqueous leaf extracts sprayed at 8 days after sowing pro-
tect chilli seedlings against Colletotrichum capsici (Sydow)
Butler and Bisby up to 35 days after sowing. During the
current study, though both aqueous and ethanol extract
of Datura metel showed significant activity (P < 0.01),
the ethanol extract was found to be more effective in
inhibiting spore germination in vitro.
Rhizome extracts of Zingiber officinale showed high
antifungal activity against Pestalotiopsis theae and
Colletotrichum camelliae. This plant is reported to pos-
sess insect growth inhibition, antifeedant, antibacterial,
and antifungal properties (Singh et al., 1991; Agarwal
et al., 2001; Martins et al., 2001). Curcuma longa also
showed significant activity that is in agreement with
findings reported in literature (Jayaprakasha et al.,
2001). In this case, the aqueous extract was more effec-
tive than the ethanol extracts. The antimicrobial proper-
ties of Azadirachta indica leaves, which showed high
antifungal activity against Pestalotiopsis theae and
Botryodiplodia theobromae, are well established (Parveen
& Alam, 1996). Literature studies on other plants
with more than 90% inhibition of spore germination
revealed that antimicrobial property of Cassia tora
(Kitanaka & Takido, 1986), Dryopteris filix-mas
(Asolkar et al., 1992), and Psidium guajava (Ali et al.,
1996) have been reported earlier. However, there are
no previous reports on antifungal activity of Leea indica,
Piper peepuloides, and Smilax zeylanica against phyto-
pathogens.
In spite of the fact that all the plants were tested for
their antifungal efficiency by using crude and concen-
trated extract, the possibility of development of effective
fungicides from plant sources to control tea-leaf dis-
eases is evident from our initial screening. Among the
four tested pathogens, all are not equally controlled
by the same plant extract. Hence, to control all the
pathogens, a bioformulation involving extracts from
more than one plant may be necessary for effective
application in the field. Thus, many of the tested plants
may be used for developing new, safer, and effective
fungicides.
 Antifungal activity of plant extracts against pathogens of tea
Acknowledgments
The authors wish to thank Prof. A.P. Das, Department
of Botany, University of North Bengal, for identification
of some plants. Financial assistance received from CSIR,
New Delhi, in the form of a project [38(0987)== EMR–II]
is gratefully acknowledged.
References
Agarwal M, Walia S, Dhingra S, Khambay BPS (2001):
Insect growth inhibition, antifeedant and antifungal
activity of compounds isolated=derived from Zingiber
officinale Roscoe (ginger) rhizomes. Pest Management
Sci 57: 289–300.
Ali MA, Shamsuzzaman M, Rahman MH, Hoque MM
(1996): Screening of different solvent extracts of the
bark of Psidium guajava for antibacterial activity.
Bangladesh J Sci Ind Res 31: 159–165.
Asha AN, Kannabiran B (2001): Effect of Datura metel leaf
extract on the enzymatic and nucleic acid changes in the
chilli seedlings infected with Colletotrichum capsici.
Indian Phytopathol 54: 373–375.
Asolkar KK, Asolkar LV, Chakre OJ (1992): Glossary of
Indian Medicinal Plants with Active Principles, Part I.
New Delhi, Publications and Information Directorate.
Chakraborty U, Dutta SK, Chakraborty BN (1991): Anti-
fungal activity of some plant extracts on phytopatho-
genic fungi. Indian Botanical Contractor 8: 107–111.
Chatterjee A, Pakrashi SC (1997): The Treatise of Indian
Medicinal Plants, Vols. 1–6. New Delhi, National
Institute for Science Communication.
Chopra RN, Nayar SL, Chopra IC (1996): Glossary of
Indian Medicinal Plants. New Delhi, National Institute
for Science Communication.
Hu K, Dong A, Sun Q, Yao X (2001): Bioactivity of 247 tra-
ditional Chinese medicines against Pyricularia oryzae.
Pharm Biol 39: 47–53.
Jayaprakasha GK, Negi PS, Anandharamakrishnan C,
Sakariah KK (2001): Chemical constituent of turmeric
oil—A byproduct from turmeric oleoresin industry
and its inhibitory activity against different fungi. Z
Naturforsch C 56: 40–44.
Kitanaka S, Takido M (1986): Studies on the constituents in
the roots of Cassia obtusifolia L. and the antimicrobial
91
activities of constituents of the roots and the seeds.
Yakugaku Zasshi 106: 302–306.
Martins AP, Salgueiro L, Goncalves MJ, da Cunha AP,
Vila R, Canigueral S, Mazzoni V, Tomi F, Casanova
J (2001): Essential oil composition and antimicrobial
activity of three Zingiberaceae from S. Tome e Pricipe.
Planta Med 67: 580–584.
Muhsin TM, Al-Zubaidy SR, Ali ET (2000): Effect of garlic
bulb extract on the growth and enzymatic activities of
rhizosphere and rhizoplane fungi. Mycopathologia
152: 143–146.
Parveen G , Alam MM (1996): Bioactivity against plant
pathogens. In: Randhawa NS, Parmer BS eds., Neem.
New Delhi, New Age International (P) Limited, pp.
192–201.
Prithiviraj B, Singh UP, Singh KP, Plank-Schumacher K
(1998): Field evaluation of ajoene, a constituent of
garlic (Allium sativum) and neemazal, a product of
neem (Azadirachta indica) for the control of powdery
mildew (Erysiphe pisi) of pea (Pisum sativum). J Plant
Dis Protection 105: 274–278.
Rana BK, Taneja V, Singh UP (1999): Antifungal activity of
an aqueous extract of leaves of garlic creeper (Adenoci-
lymma alliaceum Miers.) Pharm Biol 37: 13–16.
Rastogi RP, Mehrotra BN (1995): Compendium of Indian
Medicinal Plants, Vol. 4. New Delhi, CDRI, Lucknow
and Publications & Information Directorate.
Samuel JK, Andrews B, Jebashree HS (2000): In vitro
evaluation of the antifungal activity of Allium sativum
bulb extract against Trichophyton rubrum, a human skin
pathogen. World J Microbiol Biotechnol 16: 617–620.
Sindhan GS, Hooda I, Parashar RD (1999): Effect of some
plant extracts on the vegetative growth of root rot caus-
ing fungi. J Mycol Plant Pathol 29: 110–111.
Singh UP, Srivastava BP, Singh KP, Mishra GD (1991):
Control of powdery mildew of pea by ginger extract.
Indian Phytopathol 44: 55–59.
Srinivasan D, Nathan S, Suresh T, Perumalsamy PL (2001):
Antimicrobial activity of certain Indian medicinal
plants used in folkloric medicine. J. Ethnopharmacol
74: 217–220.
Yoshida H, Katsuzaki H, Ohta R, Ishikawa K, Fukuda H,
Fujino T, Suzuki A (1999): An organosulphur com-
pound isolated from oil-macerated garlic extract, and
its antimicrobial effect. Biosci Biotechnol Biochem 63:
588–590.