Biological Control of

Spilocaea oleagina, using

Lebanese University
Faculty of Agronomy Antagonistic Bacteria

By: Cosette Nehme 2018-2019

OUtline

Introductio M&M Results Conclusion

Olive
n in Fungal extraction Effect of Perspective
Lebanon Bacterial extracted and further
objectives extraction bacteria on the research
In vitro fungus
antagonistic test
Introduction
Introduction

 
Olive status in Lebanon
 The olive tree, Olea Europaea is one of the
oldest cultivated trees in the world.
 They are on average 150 years old.
 Area:

Source: Ministry of Agriculture

 Introduction

Causal agent
 Foliar disease
 Ascomycotina fungus
 Widely spread in all Mediterranean countries
 First reported in lebanon by Abouel-Nasr and
Boulos (1950)
 High humidity and winter conditions

Fig. 1 A, B Colony growing on MEA. C, D Conidial chains

and septate hyphae. E–J Conidiogenous cells and conidial
chains. Scale bars: C, D = 20 μm; E–J = 10 μm.
https://www.researchgate.net/publication/313576724  
Introduction

Traditional control
 Copper- containing fungicides Modern method
• Not always effective
The use of
• Resistant pathogen strains biological control
• Cu phytotoxicity agents
• Cu accumulation in the soil

Goal! Isolate potential bacterial antagonists of S.oleagina, and to

test them under in vitro conditions
Materials & Methods
 1. Fungal extraction

Segments of olive 100 μl of conidial

leaves with obvious suspension
OLS symptoms % of germination ?
OLE 100 conidia/ plate
light microscope
5 ml After 48 h
magnification : 200x
SW at 20°C
2. Bacterial extraction
Autoclaved 0.85% NaCl
containing
0.5 ml of Tween 80
At 110 rpm
and 25 °C for 10’

Sub-culturing and purifying Plating 1 ml of

48 h at 28°C NA
on another NA plate the washing
solution
3. In vitro antagonistic test

Centrifuged at 5000 rpm 25 ml nutrient

broth (NB)

 Each pellet was resuspended

in sterile 0.85% NaCl.
 Bacterial concentration was
adjusted to 1 × 109 CFU/ml
28°C
150 rpm
 3. In vitro antagonistic test

100 μl of bacterial suspension Three 20 μl droplets of

+ the suspension
100 μl of fungal conidia
+
800 μl of OLE+potato dextrose broth

3 replicate

20 ml SW agar
high H (> 85%) Germination % & Germination
20°C for 24 h inhibition rate were calculated
Results
 Effect of antagonistic bacteria
 Inhibition of S. oleagina
conidial germination and
germ tube elongation was
obvious in the presence of
the bacteria

Six bacterial isolates proved to have

a pronounced antagonistic
in vitro effect against the fungus

Fig 2: A, control; B, Pseudomona isolate Ors3;

C, Bacillus isolate Bat and D, Pseudomonas
isolate 1.1.
https://www.researchgate.net/publication/313576724  
Effect of antagonistic bacteria

 Six bacterial isolates proved to have a pronounced antagonistic in vitro effect

against the fungus

 Germination rate did not differ significantly between the different bacterial isolates

except for conidia treated with Pseudomonas isolate OLS, where the inhibition rate

(60.23%) was significantly (P < 0.05) lower than that in conidia treated with

Pseudomonas isolate ORS3 (96.95%).

 Effect of antagonistic bacteria

Germination inhibition rates of S.oleagina conidia as affected by the bacteria.

Conclusion
Conclusion

 The management of the OLS using antagonistic microorganisms has

become more important in recent years, and this can diminish the use of

chemical products. The study shows that some bacterial isolates have the

potential to inhibit conidial germination of S.oleagina in vitro.

What about the efficiency of these bacterial isolates in field conditions?!

Thank you