J. Env. Bio-Sci., 2015: Vol.

29 (2):409-411
(409) ISSN 0973-6913 (Print), ISSN 0976-3384 (On Line)

ANTIMICROBIAL PROPERTIES OF ROOT AND SHOOT EXTRACTS OF

ACHYRANTHES ASPERA L.
Mohinder Kaur* and Sheetal Rana
Department of Basic Sciences, Dr. Y S Parmar University of Horticulture and Forestry,
Nauni, Solan (Himachal Pradesh), India 173 230
[Corresponding author E-mail*: mohinderkaur9@yahoo.co.in]

Received: 29-05-2015 Accepted: 23-07-2015

Hydrophobic and hydrophilic extracts of Achyranthes aspera L. showed significant antibacterial and antifungal activities against
tested microorganisms. Chloroform and methanol extracts of roots and shoots of Achyranthes aspera L. showed significant
antibacterial activity against Klebsiella sp. while petroleum ether extract of roots showed the activity against B. substilis only.
Antifungal activity of roots was found in extracts with petroleum ether, chloroform and methanol against Fusarium sp. only.
Methanol and Aqueous extracts of shoots were also antifungal in nature against Heterobaridoin annosum and Fusarium sp.
Aqueous extracts of shoots showed weak antifungal effect against Pencillium, Phytophothora and Sclerotium sp. The present
study justifies the claimed uses of A. aspera in the traditional system of medicine to treat various infectious diseases.

Emergence of multidrug resistance in human and animal
pathogenic bacteria as well as undesirable side effects of
certain antibiotics has triggered immense interest in the search
of new antimicrobial drugs of plant origin. In recent years,
drug resistance to pathogenic bacteria has been commonly
reported from all over the world 1-2. Microorganisms and
medicinal plants have been used for several decades as rich
sources of secondary metabolites in the fields of agriculture,
industry, biotechnology and health important idiolites3-8.These
commercial products are valued in terms of billions of dollars
world wide. They have been extracted from plants for
centuries, however, their exploitations is yet to be fully
realized4. The biosynthesis of these secondary metabolites
is though largely genetically controlled and affected strongly
by the environmental influences that may be biotic or
abiotic 7-8 . As a result, there are fluctuations in the
co ncen trations and qua ntities of these sec onda ry
metabolites such as alkaloids, glycosides, volatile oils and
steroids etc 4. Developing countries like India, in particular
have an urgent need for such exploitation since they depend
to a far greater extent upon plant derived compounds. The
discovery and isolation of chemical structures of the active
antimicrobial metabolites have provided a unique starting point
for chemical modification of the biologically active compounds
in an attempt to improve their spectrum, pharamacoknetics
and human safety. Achyranthes aspera L. has valuable
medicinal properties as it is effective in curing asthma, skin
eruptions, piles, snake bite and hydrophobia9-12. It is also
reported to possess antidiabetic, antirheumatic and
antifertility properties and is also used beneficially in
abdominal tumors 9. A.aspera L. requires a detailed
investigation about the characterization of its secondary
metabolites and their biological activities.
MATERIAL AND METHODS
Preparation of plant extracts: Shade dried powdered plant
materials (100 g) of each roots and shoots of Achyranthes
aspera L. were sequentially extracted separately with
petroleum ether (40-60oC), chloroform, methanol in a soxhlet
apparatus13. Extraction with each solvent was carried out
for 18 hours. Finally, the organic solvent exhausted plant
material (roots and shoots) were extracted with cold water
for 18 hours separately. Each solvent extract was evaporated
to dryness under vacuum using a rotary evaporator and stored
at 4oC. The residues obtained were used for determination
of antim icrobial activities after m aking different
concentrations in dimethyl sulfoxide (DMSO,w/v ). 100 μ l of
each extracted sample at different concentrations (0.5-2mg/
ml) was used for estimation of antimicrobial activities by
modified well plate assay method14-15.
Well Plate assay method: Lawn of each indicator test
bacteria and fungi were made with the help of sterile cotton
swabs on nutrient and potato dextrose agar plates
respectively. Wells (8 mm) were punched on the plate with
the help of sterile cork borer. Plates were then incubated at

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 ANTIMICROBIAL PROPERTIES OF ROOT AND SHOOT EXTRACTS
37oC and 28+2oC for 24 h and 24 h-72 h, respectively, after
application of 100 ml of each concentration of plant extracts
in the well. Plates were then observed for clear zone formation
around the well. Activities were expressed in terms of diameter
(mm) of clear zone produced around the well (8 mm) at 37oC
for antibacterial and at 28+2oC for antifungal activities after 24
h and 24-72 hours of incubation, respectively.
RESULTS AND DISCUSSION
Antibacterial activity: Petroleum ether extract of roots of
Achyranthes aspera L. was antibacterial against Gram positive
Bacillus subtilis (9-10mm ), while chloroform and methanol
root extracts of roots showed significant antibacterial activity
against Gram negative Klebsilla sp. (11-14 mm). Chloroform
extracts of roots were also inhibitory against another gram
negative bacteria i.e. E. coli and Shigella sp. (Results not
shown).Chloroform and methanol extracts of shoots were also
found to active against Klebsiella (9-16 mm) . Aqueous extracts
of both root and shoot did not show any antibacterial effect
against any of the bacteria tested (Table-1).
Antifungal activity: Different concentrations (0.5-2mg/ml) of
Petroleum ether, chloroform and methanolic extracts of roots
of A. aspera L. showed antifungal effect (Table-2) against
Fusarium spp. (9-13.33 mm). Petroleum ether and methanol
extracts of roots also showed antifungal effect against Pythium
(weak) and Alternaria species (12-13mm), respectively. In case
of shoots only methanol extract of shoots showed antifungal
effect against Fusarium and Heterobaridion sp. while aqueous
extract of shoots showed weak positive effect against
Penicillium, Phytophthora and Sclerotium spp. (results not
shown). Petroleum ether extract of shoots also showed weak
antifungal effect against Phytophthora sp.
Statistically (Tables-1,2) all the extracts of roots and shoots
of plant and all the concentrations tested were significantly
different from each other. Interactions between extracts and
concentrations were significant. Statistically chloroform and
methanol extracts of roots of A. aspera showed, best antifungal
activity against Fusarium and chloroform shoot extract showed
best antibacterial activity against Klebsiella sp.
Secondary metabolites from medicinal and aromatic plants
exhibit a range of biological activities including antimicrobial,
antioxidant, insecticidal and immunosuppressive etc.
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(410)
Medicinal plants of Himalayan origin are rare and highly
useful as drug and for other purposes. Due to accelerating
process of deforestation and soil erosion, many species
are being lost which might have yielded useful products to
mankind and profit to the industry. Antimicrobial effects of
hydrophobic and hydrophilic extracts of roots and shoots
of H. candicans were tested against Gram positive and
Gram negative bacterial species and fungal species.
The variation in antimicrobial activity may be due to the
difference in the chemical nature of cell walls and cell
membranes of each bacteria. The information regarding
antimicrobial and pharmacological activity about this plant is
very scanty. The effectiveness of A. aspera L. to control
infectious diseases may be due to presence of unknown
antimicrobial secondary metabolites. Only few preliminary
investigations showed that shoots of A. aspera possessed
essential oil and new long chain alcohol and their essential
oil exhibited antifungal activity against Aspergillus corneus.
Earlier workers have shown that antibacterial effect of alcoholic
extracts against B. subtilis, Staphylococcus aureus,
Pseudomonas aeruginosa and Shigella dysentriae is due to
presence of alkaloids AM-I and AM-II10-12. Some workers also
have also shown the antibacterial effect of alcohol root extract
of Achyranthes bidentata against E. coli, S. aureus and
Pseudomonas sp. But have not purified and identified the
effective component. In our studies antibacterial and antifungal
effect of different extracts may also be due to presence of
alkaloids, long chain of alcohol or essential oil or may be due
to some another unidentified bioactive secondary metabolites
with antimicrobial activities. But the plant still requires detailed
investigation about the characterization of these secondary
metabolites. The emergence of multidrug resistance to
microorganisms has also necessitated a search for new
antimicrobial substances from other sources including plants
Different extracts from traditional medicinal plants have been
tested and some natural products have been approved as
new antimicrobial drugs but there is still need to identify more
substances active towards pathogens16-18. Present finding
support the applicability of A. aspera L. in traditional system
for it’s claimed uses. Further work is necessary for isolation
and purification of compound in root and shoot extracts which
will allow the scientific communities to recommend the
utilization as an accessible alternative to synthetic antibiotics.
 (411) KAUR AND RANA

Table-1. Antibacterial activity of root & shoot extract of Achyranthes aspera L. by well plate method.

CD = 0.05; Conc. 0.16; Indicator test bacteria 0.18; Conc. x Indicator test bacteria 0.26
*Antibacterial activity expressed in terms of mm, diameter of clear zone (mm dia.) produced around the well (8 mm) by
100 µl of plant extract at 37 oC for 24 hours.
Table-2. Antifungal activity of root & shoot extract of Achyranthes aspera L. by well plate method.

Effect CD0.05
Concentration = 0.19
Indicator test bacteria = 0.23
Conc. x Indicator test bacteria = 0.46
Antifungal activity expressed in terms of mm, diameter of clear zone (mm dia.) produced around the well (8 mm) by
100 µl of plant extract at 28+2oC for 24-72 hours.

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