ISSN: 0974-6943

Research Article
Bioactivity of Andrographis paniculata Against Selected Phytopathogens
Varaprasad Bobbarala*, P. Koteswara Rao**, G. Srinivasa Rao* and D. Aryamithra*
*For U Biosciences, A/4A, Park lane Residency, East point colony, Visakhapatnam, A. P-530017, India.
**Department of Biochemistry, Andhra University, Visakhapatnam, A.P .530003
For correspondence:Varaprasad Bobbarala ,U Biosciences, A/4A, Park lane Residency, East point colony,
Visakhapatnam, A. P-530017, India.
E-mail:varaprasadphd@rediffmail.com
Received on:19-08-2008; Accepted on :08-01-2009

ABSTRACT
The objective of this study is to determine the potential antimicrobial activity of three organic solvent viz., Hexane, chloroform
and methanolic extracts of Andrographis paniculata, was investigated for their antimicrobial activity against phytopathogenic
bacteria and fungi. The bacteria include Erwinia caratovora, Pseudomonas marginales, Pseudomonas syringae,
Pseudomonas aeruginosa and Xanthomonas compestris and fungal pathogens include Acremonium strictum,Alternaria
alternate, Aspergillus flavus, Bipolaris bicolor, Cladosporium herbarum, Curvularia lunata, Fusarium oxysporum,
pencillium expansum, Rhizoctonia solani, Tiarosporella phaseolina and Ustilago maydis using the Well diffusion method.
Organic solvent extracts of Andrographis paniculata have shown broad spectrum of antibacterial as well as antifungal
activity against tested organisms. The growths of all bacterial and fungal pathogens were highly inhibited by Methanolic
extracts of Andrographis paniculata than chloroform and hexane extracts respectively. The methanolic extracts inhibited
the growth of 95% organisms tested, followed by chloroform extracts inhibited 80%, Hexane extracts 65% growth of
tested organisms.
Key words: Andrographis paniculata, Organic solvent extraction, Antimicrobial assay, Growth inhibition, Phytopatho-
gens.
INTRODUCTION
Microbial diseases are responsible for widespread mortality simplex virus 1(HSV-1) dueto ent-labdene diterpenes iso-
in the world. For centuries plants have been used throughout lated from Andrographis paniculata was also reported [8].
the world as drugs and remedies for various diseases. Many Asmah Rahmat et al., in 2006 [9] reported the antidiabetic
efforts have been made to discover new antimicrobial com- effect of crude extract of Andrographis paniculata leaves
pounds from various kinds of sources such as soil microor- on normal and alloxan induced hyperglycemic rats. The car-
ganisms, animals and plants [1]. Decreased efficacy of anti- diovascular effects of 14-deoxy-11,12-didehydro
biotics and resistance of pathogens to antibiotics has neces- andrographolide (AP3) of Andrographis paniculata extracts
sitated development of new alternatives [2]. Making antimi- were studied by Yoopan et al., in 2007 [10]. They reported
crobial drug therapy effective, safe and affordable has been that AP3 was most potent compound for inducing vaso re-
the focus of interest during recent years [3] there are severallaxation and decreasing heart rate. Based on the above infor-
reports on antimicrobial activity of different herbal extracts mation present investigation has been carried out to evaluate
[4]. Andrographis paniculata is one of the important herbs antimicrobial activity of Andrographis paniculata against plant
comprising several medicinal properties. Andrographis bacterial and fungal pathogens.
paniculata is belonging to the family Acanthaceae. MATERIALS AND METHODS
Andrographolide is the major constituent extracted from the Andrographis paniculata, (AP), also known com-
leaves of the plant which is a bicyclic diterpenoid lactone. monly as “King of Bitters,” is a member of the plant family
Andrographis paniculata has been found to be an Acanthaceae, and has been used for centuries in Asia to treat
effective antibiotic, antiviral, anti-parasitic and immune sys-gastro intestinal tract and upper respiratory infections, fever,
tem stimulant. Hepatoprotective activity of A. paniculata herpes, sore throat, and a variety of other chronic and infec-
against carbontetrachloride was studied [5]. tious diseases. It is found in the Indian Pharmacopoeia and
Immunostimulating agents present in Andrographis is the prominent in at least 26 Ayurvedic formulas; whereas in
panivulata were studied by Puri et al [6]. According to the Traditional Chinese Medicine (TCM), Andrographis is an
studies of Rajgopal et al, in 2003 [7] andrographolide is an important “cold property” herb: it is used to rid the body of
interesting pharmacophore with anticancer and heat, as in fevers, and to dispel toxins from the body. Three
Immunomodulatory activities.Viricidal activity against herpes different organic solvents such as Hexane, chloroform and
Journal of Pharmacy Research Vol.2.Issue 3.March 2009 480
 ISSN: 0974-6943

Table: 1 Andrographis paniculata antimicrobial activity on phytopathogens

Microorganism Hexane Chloroform Methanol

100mg/ml 250 mg/ml 100mg/ml 250 mg/ml 100mg/ml 250 mg/ml
Pseudomonas marginales 10mm 13mm 12mm 20mm
15mm 20mm
Pseudomonas syrengii — 13mm 12mm 15mm 14mm 15mm
Erwinia caratovora — — 14mm 15mm
13mm 14mm
Pseudomonas aeuriginosa — — — —
10mm 19mm
Xanthomonas campestris — — — 15mm
— 14mm
Acrimonium strictum 9mm — 16mm —
10mm —
Alternaria alternata — 10mm 12mm 14mm 17mm 18mm
Aspergillus flavus 11mm — 15mm —
12 —
Bipolaris bicolor 8mm 10mm 13mm 16mm
10mm 10mm
Cladosporium herbarum — 10mm — 12mm
10mm 10mm
Curvularia lunata 8mm 8mm 12mm 13mm
15mm 15mm
Fusarium oxysporum — 13mm 18mm 20mm 13mm 14mm
Pencillium expansem 11mm — 14mm —
12mm —
Rhizoctonia solani — — 15mm 15mm
10mm 11mm
Tiarosporella phaseolina 11mm — 15mm —
8mm —
Ustilago maydis — — 13mm 13mm
8mm 8mm

methanol were used respectively depending on their increas-
ing order of polarity for extraction of bioactive principles.
Andrographis paniculata was collected from bo-
tanical garden in Andhra University, Visakhapatnam. Then
plant material was dried under shade, powdered using bender
and stored in airtight bottles. The powdered material (150g)
was extracted with organic solvents viz., Hexane chloroform
and methanol respectively according to their increasing order
of polarity. With three different solvents, Andrographis
paniculata was extracted by hot extraction process using
soxhlet apparatus followed by condensation under vaccum
(50°c) and then dried in vacuum desicator. The extracts of
Andrographis paniculata autoclaved and stored at 4°c.These
extracts can be used for further screening of antimicrobial
activity against tested organisms.
Bacterial and fungal strains causing infectious diseases
in plants were used in current study. Among the plant patho-
gens Bacterial ones include Erwinia caratovora , Pseudomo-
nas marginales, P. syringe, P .aeuriginosa, Xanthomonas
campestris and Fungal pathogens include Acrimonium
strinctum, Alternaria alternate, Aspergillus
flavus,Bipolaris bicolor,Cladosporium herbarum,
Curvularia lunata, Fusarium oxysporum, Pencillium
expansum, Rhictonia solani, Tiarosporella phaseolina,
Ustilago maydis. The bacteria were grown in the nutrient
broth at 37°C and maintained on nutrient agar slants at 4°C.
Similarly the fungal strains were grown in potato dextrose
broth at 37°C and maintained on potato dextrose agar slants
at 4°C. Nutrient agar medium was used to study the antibac-
terial activity where as potato dextrose agar was used to study various sources, such as medicinal plants. The screening of
the antifungal activity of crude extract of Andrographis
Journal of Pharmacy Research Vol.2.Issue 3 March 2009
paniculata. Pure cultures of bacterial and fungal pathogens
were removed from agar slants and transferred to nutrient
broth (bacteria) and Potato dextrose broth (Fungi) and incu-
bated at 37°c for 24-48h.The turbidity was adjusted to that
of standard level by adding sterile nutrient and PD broth.
Antimicrobial screening:
The hexane, chloroform and methanol extracts of
Andrographis paniculata extracts were screened for anti-
microbial activity by agar well diffusion method [11] with Cork
borer of size 6.0mm. For all bacterial and fungal strains, over-
night cultures grown in nutrient and potato dextrose broth
were used for inoculation of the agar plates. An aliquot (0.02ml)
of inoculum was introduced to molten and cooled at 45°C
nutrient or potato dextrose agar and placed on petriplate by
pour plate technique. Then appropriate Wells were made on
respective agar plate by using cork borer. In agar well diffu-
sion method 0.05ml of Hexane, chloroform and methanol
extracts were introduced to their respective wells following
an incubation period of 24to48h at 37°C. Antimicrobial ac-
tivity was evaluated by qualifying zones of inhibition of bacte-
rial growth surrounding the plant extracts. The entire antimi-
crobial assay was carried out under strict aseptic conditions.
The result of screening is shown in the table 1.
RESULTS AND DISCUSSION
The antimicrobial screening of three different organic
solvent extracts of Andrographis paniculata are presented
in table 1. The undesirable side effect of certain antibiotics,
and the emergence of previously uncommon infections, has
forced scientists to look for new antimicrobial substances from
plant extracts and plant products for antimicrobial activity has
481
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tial for use as antimicrobials to human population.
Source of support: Nil, Conflict of interest: None Declared
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