Presentation 01

Exploration of endophytic
microorganisms from selected

medicinal plants and their control
potential to multi drug resistant
pathogens
Author: Arun Kumar G, Robert Antony A, V. Rajesh
Kannan
Journal of Medicinal Plants Studies 2015; 3(2):
49-57
Reviewed by: Khan Naushad M.

Seat no. 3018
Semester 4th
M. Sc Botany
Department of Bioscience, VNSGU
Introduction
• The World Health Organization reports that 25% of annual global deaths are
caused by microbes.
• Methicillin-resistant Staphylococcus aureus (MRSA) strains are multidrug-
resistant and pose a significant public health threat.
• New antibiotics are necessary due to the rise of resistant pathogens and the
toxicity of current drugs.
• Endophytes, microbes that live within plant tissue without causing harm, have
been studied for their potential in producing bioactive compounds with medicinal
properties.
• These compounds may play a role in plant defense and novel drug discovery.
• Research on endophytes has led to the discovery of numerous natural products,
including antibiotics, with significant antimicrobial properties.
• Despite the vast potential of endophytes, much of their biodiversity remains
unexplored, highlighting the importance of further research in this area to uncover
Material and Methods
Sample Collection
Isolation of Endophytic Microorganism
Fungal and Bacterial Isolation
Screening of Antimicrobial Compounds
Extraction of Antimicrobial Compounds from the
Evaluation of antimicrobial activity
Identification by 16s rRNA sequencing
GCMS Analysis
Sample Collection
1 Terminalis arjuna
3 Azadirachta
2 Catharanthus roseus indica
Isolation of Endophytes
Leaves and
Surface
stem were
sterilized leaf
washed and
Sample and stems
immersed in
plant cut placed in
70% alcohol
into small Potato
followed by
pieces Dextrose agar
sodium
medium and
hypochlorite
incubated
for few sec.
Fungal and Bacterial Cultivation
Fungal Culture Bacterial Culture
Agar blocks placed in Potato Bacteria is inoculated in

dextrose broth Nutrient broth
Incubated at 25± 1 for 3 weeks Incubated at 30 for 3

with periodically shaking at days
150 rpm
Culture centrifuge at
Culture filtered to remove the 8000 rpm and collect
mycelial mats supernatant
Extraction of Antimicrobial Compoun
creening of Antimicrobial Compound
from the Bacterial Endophytes
Cross Streaked method
Single pure bacterial endophytes
were inoculated in nutrient broth
culture were streak on one edge of
the Nutrient agar plate Incubated in a rotary shaker at 30C,
120 rpm for 5 days
Human bacterial pathogen were Centrifuge at 8000 rpm and pellet
horizontally streaked from other was collected
edges
5 different polar and nonpolar
Incubated at 37 for 24 – 48 hours solvents were added and mixed
Antimicrobial compound from
Zone of inhibition was measured supernatant was evaporated
Evaluation of Antimicrobial Activity
Well or Cup agar diffusion methods
Cup agar were filled with bacterial crude extract and swapped by
• Staphylococcus aureus
• klebsiella pneumonia
• Enterococcus faecalis
• Pseudomonas aeruginosa
• Candida albicans
Identification by 16s rRNA
sequencing
Genomic Agarose Gel Amplificatio
Sequenci
Nucleotide
ng of 16s
DNA Electrophor n of 16s Sequence
rRNA
Isolation esis rRNA gene Analysis
gene
Result
Isolation of Endophytes
Screening for Antimicrobial Activity
Evaluation of Antimicrobial Activity
Methanol Extract to Control the S. Ethanol Extract to Control the S.

Phylogenetic Analysis
GC-MS Analysis
GC-MS Analysis
Discussion
• In this study, endophytic organisms were isolated from three
medicinal plants: Azadirachta indica, Terminalia arjuna, and
Catharanthus roseus.
• A total of 14 fungi and 7 bacteria were isolated from these plants,
and the bacterial isolates were screened for antimicrobial activity
against multidrug-resistant pathogens.
• Among the isolated organisms, one bacterial isolate demonstrated
effective antimicrobial activity against Methicillin-resistant
Staphylococcus aureus.
• Additionally, endophytic fungi were found to produce bioactive
secondary metabolites with antibacterial properties.
• Chemical analysis identified several compounds responsible for the
antimicrobial activity of the extracts.
• Moreover, sequencing of bacterial isolates revealed their identity as
Bacillus cereus and Bacillus subtilis.
My Review
Positive Negative
• Bacterial isolates showed • While positive results are showcased
antimicrobial activity against drug- in the images, the lack of negative or
resistant pathogens, including control data limits the
MRSA. Endophytic fungi produced comprehensiveness of the study,
antibacterial compounds. leaving questions about the overall
• The paper offers a detailed validity of the findings.
description of the experimental • The absence of an incubator in the
procedure, enhancing transparency experimental setup raises concern
and reproductivity. about the reliability of the results, as
• All the measurements are clearly temperature control is crucial for
documented. many biological processes.
• The majority of work was
conducted in lab, demonstrating a
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Exploration of endophytic

microorganisms from selected

Reviewed by: Khan Naushad M.

Isolation of Endophytic Microorganism

Fungal and Bacterial Isolation

Screening of Antimicrobial Compounds

Extraction of Antimicrobial Compounds from the

Evaluation of antimicrobial activity

Identification by 16s rRNA sequencing

Fungal Culture Bacterial Culture

Agar blocks placed in Potato Bacteria is inoculated in

Incubated at 25± 1 for 3 weeks Incubated at 30 for 3

Evaluation of Antimicrobial Activity

Methanol Extract to Control the S. Ethanol Extract to Control the S.

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