1st International Electronic Conference On Plant Science

1 to 15 December 2020

Biocontrol activity of actinomycetes strains against fungal and bacterial

pathogens of Solanum lycopersicum L. and Daucus carota L.: in vitro and
in planta antagonistic activity
Rihab Djebaili1, 2,*, Marika Pellegrini2, Matteo Bernardi2, Maria Smati1, Mahmoud Kitouni1, and Maddalena Del Gallo2

2020/202
 Introduction

-Plants are affected by different biotic and abiotic stresses due to climate change.

-Increasing in temperature make plants susceptible to pathogens attack.

-Tomatoes and carrots are important crops most consumed worldwide for their nutritional values, these
vegetable crops are attacked by several pathogens which limit their growth and productivity.

-Several rhizospheric microorganisms have the ability to promote plant growth, and have antagonistic
properties against pathogens attack, and play a role in disease control.

-Plant growth-promoting rhizobacteria (PGPR) have a beneficial effects on plants by direct and indirect
mechanisms; phosphate solubilization, siderophores production, nitrogen fixation, the activity of 1-
aminocyclopropane-1-carboxylate deaminase enzyme, fixation of atmospheric nitrogen, phytohormones
and plant growth regulators production, protection against phytopathogens by systemic resistance (SRI)
induction. (Nawaz et al., 2020; Orio et al., 2016; Pagnani et al., 2018)
• Actinomycetes are a group of filamentous bacteria widely distributed in soil, GRAM
positive, with a high percentage of GC, they are resistant microorganisms, can sporulate
when environmental conditions become unfavorable.

• They have the capacity to produce various metabolites, hydrolytic enzymes and to degrade
recalcifying materials.

• The genus Streptomyces is the most reactive, it is responsible of a wide range of antibiotic
production. It constitutes an important group of microorganisms due to their antimicrobial
power, dominance in soils and their promotion of plant growth.

• They have the ability to inhibit a large spectrum of pathogenic bacteria, and several traits
promoting plant growth and suppress plant diseases.

(Gowdar et al., 2018)

• Stimulation of plant growth includes hormones, and plant growth regulators
production such as indole acetic acid (IAA) and cytokinin which are involved in
root development, exudation and plant growth.

• these bio-stimulators and biocontrol agents represent a substitute for harmful

chemical pesticides for achieving the quest for sustainability in agriculture.

(Gowdar et al., 2018)

 Experimental part

• In vitro antagonistic activity of diffusible and volatile compounds was carried out by dual culture
method against Fusarium oxysporum f. sp. radicis-lycopersici (FORL), and Rhizoctonia solani
(RHS).

• PGPB-fungus interaction zone were analyzed by SEM microscopy.

• The pathogenic bacteria tested were: Pseudomonas syringae, Pseudomonas corrugata,

Pseudomonas syringae pv. actinidiae, Pectobacterium carotovorum subsp. Carotovorum.

• The inhibition percentages were calculated after incubation until the complete growth of the
control plate.
 Results

Strains H12 (Streptomyces albidoflavus), and H14 (Nocardiopsis aegyptica) showed good in
vitro antagonistic activity by diffusible and volatiles compounds against both pathogens
(inhibition percentage up to 85%).

Figure1. Antagonistic activity by diffusible and volatile compounds by dual culture

method. (A) diffusible compounds. (B) volatiles compounds. (C) normal growth of
fungul strain.
Figure 2. SEM micrographs of FORL and RHS hyphae. The panels show the fungal hyphal branching
following a normal growth in FORL and RHS control plates (A and B, respectively). In the presence
of H12 and H14 consortium, the hyphal structures change in the interaction zones of both FORL and
RHS with the PGPB (C and D, respectively).
• Strains H12, and H14 showed good antibacterial activity against Pseudomonas

corrugate, and Pectobacterium carotovorum (inhibition diameter ˃10mm).

• The CFCs of combined strains were investigated for cell-free supernatant (CFS)

Minimal Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration

(MBC).

The CFSs (up to 0.8 %) was effective to inhibit the growth of both FORL, and RHS
(Wayne, 2011)
fungi and most of the bacterial pathogen growth.
• In planta antagonistic activity of H12 and H14 consortium was assessed on S.
lycopersicum, and D. carota in pre-emergence.

• The experiment was organized as follows: (i) CNT (seeds without

PGPB/infection), PGPB (seeds with PGPB/no infection), PGPB+INF (seeds
with PGPB/infection), INF (seeds with infection/no PGPB).

• The induced protection was assessed by estimation of plant survival, morpho-

biochemical parameters, damages and chlorophyll contents.

(Pellegrini et al., 2020)

Table1. Results of in planta pre-emergence (PRE) antagonistic activity of bacterial consortium (PGPR) against fungul
and bacterial strains.
S. lycopersicum D. carota

Parameters CNT PGPR PGPR+F F CNT PGPR PGPR+F F

Germination 80 100 73 53 88 100 48 29,3

damages 0 0 4 4 0 0 48 3,6

Plant height 6,5 10,9 7 4 4,5 5,4 4,6 3,4

Root length 1,1 2,8 1,8 1,2 1,7 1,9 1,4 1,04

leaves 2,6 5,1 3,7 2,5 2,0 3,0 2,3 1,6

Chlo (a+b) 3,39 9,42 4,46 1,6 4,93 15,54 6,1 2,4
• Inoculation with the consortium (PGPB) improved development and growth of both
tomato and carrot plants compared to the control (better germination rates, morpho-
physiological characters, and chlorophyll content).

• Infection decreased germination rates, growth parameters, and total chlorophyll

content, and caused damages with extension up to ˃ 20mm in uninoculated plants
(INF). Treatment with the consortium improved germination in infected plants
(PGPB+INF) up to 15-54% for tomato, and up to 30-100% for carrots.

• The presence of PGPB also alleviated infection symptoms; PGPB+INF plants

showed less damages, and better chlorophyll content than the control (p <0.05).
 Conclusions and perspectives

• Plant growth promoting traits of actinomycetes strains act indirectly in the control of plant diseases.
• The use of combined bacteria is a strategy for plants protection against pathogens attack.
• Actinobacteria are well known for their ability to produce various bioactive compounds, they are biological
agents for their antagonistic activities, and plants protection against several soil borne pathogens.
• The actionmycetes strains investigated in this study also have different plant growth-promoting traits. These
traits enhance plant physiological status and offer an additional advantage to their use as biological control
agents for sustainable agriculture.
• Further experiments are needed to determine their effectiveness on other plants, against other pathogens and
under different cultivation conditions. However, these preliminary results underline that actinomycetes, and in
particular Streptomyces and Nocardiopsis genera, can be biological alternatives for plants disease management.

(Djebaili et al., 2020; Pellegrini et al., 2020)