1.

(a) ISOLATION AND CHARACTERIZATION OFENDOPHYTIC

BACTERIA FROM
PLANTS AND STUDY ON THEIR BIACTIVITIES
AND APPLICATION.
(b) Environment
(c) Applied research
2.
(a) METERIALS AND METHODS
1. Collection of plant materials
Healthy plant materials of Oscimum sanctum, Pisum
sativum, Orysa sativa are collected.
2. Sterilization of plant materials (Santos et al., 2003)
Surface sterilization was performed by sequentially
rinsing the plant materials with 70% ethanol for 30 minutes,
then with 0.01% mercuric chloride for 5 minutes followed by
0.5% sodium hypochloride for 2-3 minutes and finally with
sterile distilled water for 2-3 times. Plant material was then
dried in between the folds of the sterile filter papers.
3. Isolation of the endophytic bacteria
Small pieces of surface sterilized plant materials
inoculated into Luria Bertani agar medium and Amphotericin B
which act as antifungal agents.
4. Identification
Cultural, staining, biochemical and molecular
characteristics can be studied to identify the isolates.
5. Maintenance of endophytes
Purified endophytes were maintained at 4 0c.

6. Screening of isolates
a. Indole acetic acid production
b. Hydrogen cyanide production
c. Phosphate solubilisation
d. Production of ammonia
e. Antibacterial activity
f. Antifungal activity etc.
7. Molecular identification of endophytic colonizing bacteria
The isolates were then subjected to molecular
identification analysed for their identification.
a.
b.
c.
d.

Extraction of genomic DNA

Polymerase chain reaction (PCR) amplification
Nucleotide sequence determination
Phylogenetic analysis

(b) OBJECTIVE
To isolate, identify and characterize endophytic
bacteria from plants.
To detect the bioactivities of these isolates and study
its application.
3. APPLICATIONS
Endophytic bacteria show important role in biological
control of plant pathogens and plant growth promotion.
Endophytes have the potential application in
phytoremediation , bioremediation of hazardous
chemicals , natural products and sustainable
agriculture.

4. ABSTRACT
The endophytic bacteria colonize healthy plants without
causing apparent disease o damage to the host plant. The
present study aims that the isolation, characterization,
bioactivities of endophytic bacteria from Oscimum sanctum,
Pisum sativa, Orysa sativa and their application in various
fields.