Biological Control 86 (2015) 14–19

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Biological Control
journal homepage: www.elsevier.com/locate/ybcon

Evaluation of bacterial antagonists of Ralstonia solanacearum, causal

agent of bacterial wilt of potato
Zeinab Kheirandish, Behrouz Harighi ⇑
Department of Plant Protection, University of Kurdistan, Sanandaj, Iran

h i g h l i g h t s g r a p h i c a l a b s t r a c t

 Pseudomonas and Paenibacillus strains

showed plant growth promotion
effects on potato plants.
 Paenibacillus strain showed
antagonistic potential against
Ralstonia solanacearum.
 Pseudomonas strains showed
biocontrol activity against Ralstonia
solanacearum.

a r t i c l e i n f o a b s t r a c t

Article history: Bacterial wilt of potato caused by Ralstonia solanacearum is one of the most destructive diseases in
Received 20 April 2014 Kurdistan province, Iran. The objective of the present study was to evaluate antagonistic effects of some rhi-
Accepted 25 March 2015 zobacteria isolated from the rhizosphere of potato plants against R. solanacearum, the agent of potato
Available online 6 April 2015
bacterial wilt. A total of 52 rhizobacteria were isolated and screened for in vitro antagonistic activity against
R. solanacearum. Seven isolates with inhibiting effects of the pathogen were identified by phenotypic prop-
Keywords: erties and partial sequencing of 16s rRNA as Pseudomonas fluorescens Pf11, P. fluorescens Pf16, Pseudomonas
Ralstonia solanacearum
putida Pp17, Paenibacillus sp. Pb28 and Enterobacter sp. En38, Pseudomonas fluorescens Pp23 and Serratia sp.
Biocontrol
Bacterial antagonist
Se40. Strains Pf11, Pf16, Pp17 and Pb28 significantly inhibited the growth of the pathogen. Strains En38,
Iran Pp23 and Se40 showed a moderate or weak inhibition. During greenhouse study, strains were evaluated
for their effects in reducing of disease and increasing biomass of potato plants. In according to greenhouse
experiment results, isolates Pb28, Pp17 and Pf11significantly reduced disease by 55.56%, 51.50% and
38.58%, respectively. In addition, plant biomass significantly increased in plants treated with Pb28, Pp17,
Pf11 and Pf16, compared to the control. Therefore, this study shows that these four strains have potential
to be used as biocontrol agents against R. solanacearum. To confirm their effectiveness as commercial
biocontrol agent, it is necessary to evaluate their efficiency in the field conditions in the next studies.
Ó 2015 Elsevier Inc. All rights reserved.

1. Introduction a great number of diseases, among which wilt caused by

Potato (Solanum tuberosum L.) is one of the most important
crops growing in Iran, and about 173000 hectares of agricultural
lands are devoted to potato cultivation. The crop is susceptible to
⇑ Corresponding author. Fax: +98 8733620553.
E-mail address: BHarighi@uok.ac.ir (B. Harighi).
Ralstonia solanacearum (Smith) (Yabuuchi et al., 1995) is known
as one of the most destructive agents in temperate regions of
Iran (Azadvar and Rahimian 2006; Maghooli et al., 2004;
Irandoust et al., 2008). Because of the widespread geographic
distribution, high survival properties and unusual wide host range
of the pathogen, control methods such as crop rotation, field
sanitation and use of resistant varieties provide limited success.

http://dx.doi.org/10.1016/j.biocontrol.2015.03.007
1049-9644/Ó 2015 Elsevier Inc. All rights reserved.
 Z. Kheirandish, B. Harighi / Biological Control 86 (2015) 14–19
R. solanacearum can survive in latently infected potato tubers,
perennial weed hosts and in soil for more than one year. Thus, crop
rotation with non-host plants should be done for several years
(Van Elsas et al., 2000). Chemical control of the disease is usually
ineffective and soil fumigation has slight or no effect (Murakoshi
and Takahashi, 1984). In some plants such as tomato, application
of the chemical elicitor acibenzolar-S-methyl can reduce bacterial
wilt incidence in cultivars with moderate resistance, but is ineffec-
tive in susceptible cultivars (Pradhanang et al., 2005). Several resis-
tant cultivars are available but strain diversity of the pathogen
turns resistance unstable in different countries (Hartman and
Elphinstone, 1994; Kim-Lee et al., 2005). Therefore, biological con-
trol is an alternative method to resolve some of these difficulties.
Several researches have been carried out to find biocontrol agents
to reduce bacterial wilt severity. Plant growth-promoting rhi-
zobacteria (PGPR) colonize root surfaces and are present in the rhi-
zosphere of many plant species (Kloepper et al., 1999). These
beneficial microorganisms can increase plant growth and also pro-
tect plant from infection of plant pathogens. The mechanisms of
disease control by beneficial bacteria include the production of
antibiotics, hydrogen cyanide, siderophores and competition for
nutrients (Van Elsas et al., 2000). The objectives of this research
was to investigate the antagonistic potential of rhizobacteria iso-
lated from potato growing fields against R. solanacearum under lab-
oratory and greenhouse conditions, and evaluate the effect of
antagonists on potato plant biomass.
2. Material and methods
2.1. Isolation, characterization and identification of bacterial pathogen
Potato plants with typical symptoms of bacterial wilt were col-
lected from potato fields in various locations of Kurdistan province,
western Iran. Small pieces of tuber showing brown rot symptoms
were excised, after surface sterilization with 0.25% sodium
hypochlorite for 30s, rinsed in sterile distilled water. Each piece
was macerated in 1 ml sterile water and the resulting suspension
was streaked onto casamino acid-peptone-glucose + 2,3,5-triph-
enyl tetrazolium chloride (CPG + TZC) medium (Kelman, 1954).
The Plates were incubated at 28 °C for 72 h. After that, the colonies
typical of R. solanacearum were streaked on the same medium to
get pure colonies. Phenotypic properties of the isolates were
characterized using standard procedures (Hayward, 1964;
Yabuuchi et al., 2005). Identification of bacterial isolates was fur-
ther confirmed by polymerase chain reaction (PCR) using species-
specific 759/760 primers according to a method previously
described (Opina et al., 1997).
2.2. Isolation, characterization and identification of bacterial
antagonists
The potential bacterial antagonists were isolated from the rhi-
zosphere of healthy potato plants from different locations of
Kurdistan province during July to September 2011 and 2012. Soil
samples were collected from root zone of potato plants, placed in
plastic bags and kept in refrigerator until used. One gram of each
soil sample was suspended in 100 ml sterile–distilled water, serial
dilutions of 101–105 were prepared and 0.1 ml of each dilution
was inoculated in King’s Medium B Agar (KMB) plates (King
et al., 1954). The plates were incubated at 28 °C for 48–72 h and
observed under UV transilluminator. Colonies showing fluorescent
pigment and some non-fluorescent colonies were subcultured in
KMB to get pure cultures. Biochemical and physiological properties
of bacterial antagonists such as: gram reaction, oxidase activity,
levan production, arginine dihydrolase, gelatin hydrolysis, nitrate
15
reduction, growth at different temperatures, etc. were character-
ized according to standard methods (Schaad et al., 2001) and their
identification was further confirmed by partial nucleotide
sequencing of 16S rRNA gene using PCR with rP1/fD2 primers
(Weisburg et al., 1991). The PCR products were sequenced using
an ABI3730XL DNA sequencer (Applied Biosystems). The 16s
rRNA sequences obtained in this study were deposited in
GenBank under the accession numbers listed in Table 1. All
sequences were compared with NCBI nucleotide sequence data-
base using blast program. Nucleotide sequences were aligned using
the ClustalW available in MEGA 5.2 program (Tamura et al., 2011).
A phylogenetic tree was constructed by the neighbour-joining
method (bootstrap analysis with 1000 replicates were conducted)
with the same program.
2.3. In vitro assay of rhizosphere bacteria
2.3.1. Screening of rhizobacteria against bacterial pathogen
Three hundreds lL of the pathogen suspension (about
2  108 CFU/mL) was poured into the plates and maintained in a
laminar flow hood for 5 min. After which, paper disc (about
10 mm) was immersed in each suspension of rhizobacteria spec-
trophotometrically adjusted (absorbance at 600 nm) to concentration
of about 108 CFU/mL and was spotted at the pathogen-inoculated
plates. The plates were incubated at 28 °C for 48–72 h and the width
of the inhibition zones was measured. Sterile water spotted in the
plates with the pathogen was used as control.
2.3.2. Protease, hydrogen cyanide and siderophore production by
rhizobacteria
Bacterial isolates were tested for protease production using skim
milk (SKM) agar medium (Chakravarty and Kalita, 2012). The iso-
lates were spot inoculated into SKM medium and plates were incu-
bated at 28 °C for 48 h. Production of clear halo zone around the
bacterial colony was taken as evidence of protease production.
Production of Hydrogen cyanide (HCN) by bacteria was tested
according to the method described by Alstrom (1987). One hun-
dred lL of bacterial suspension was streaked onto plates contain-
ing nutrient agar medium. Plates sealed with parafilm were
incubated in an inverted position at 26–28 °C with picric acid indi-
cator papers placed inside the lids.
Siderophore production test was done according to a method
previously described (Kraus and Lopper, 1990). Rhizosphere bac-
teria were spotted on King B medium containing 1000 lM FeCl3
and incubated at 26–28 °C for 48 h. After that, bacterial colonies
were wiped off from the plates using sterile cotton swap.
Subsequently, pathogenic bacterial suspensions were spread on
media and incubated at 26–28 °C for 24 h. Inhibition of pathogenic
bacteria around spots indicated siderophore production.
2.4. Greenhouse experiment
A substrate mixture including field soil:sand:compost
(1.25:1.25:0.5) was prepared and autoclaved at 121 °C for 2 h.
Pots were sterilized with 1% sodium hypochlorite and three kilo-
gram of the substrate was located into each pot. Pieces of potato
tuber (var. Fontane) were surface sterilized, air-dried and dipped
into aqueous suspension of antagonist (2  108 CFU/mL) for 1 h,
then planted in pots. After 24 h, 200 ll of each pathogenic bacterial
suspension (108 CFU/mL) was added to pots.
Tubers dipped in sterile distilled water and pathogenic bacterial
suspension was planted as negative and positive controls, respec-
tively. The pots were placed in a greenhouse (26–28 °C, RH30%,
12 h/ 12 h photoperiod). Effects of bacterial antagonists on plant
fresh weight of shoots and roots were measured after 60 days.
Plants then cut above ground, shoots and roots were oven-dried
16 Z. Kheirandish, B. Harighi / Biological Control 86 (2015) 14–19

Fig. 1. Agarose gel electrophoresis of PCR product using R. solanacearum species-specific primer pairs 759/760. Lane1-6: R. solanacearum isolates, lane M: 1Kb DNA ladder.

Fig. 2. The phylogenetic tree shows the position of antagonistic strains among the type strains of the species of the genera Enterobacter, Paenobacillus, Pseudomonas and
Serratia.

at 75 °C for 48 h and dry weight was measured. The relative of dis- %reduction of disease
ease incidence and biocontrol efficacy was calculated using the fal- ¼ ½ðdisease incidence in control pots  disease incidence in
lowing formula (Algam et al., 2013; Seleim et al., 2011):
antagonist treated potsÞ=disease incidence in control  100:
%disease incidence ¼ ðnumber of wilted leaves per pot=
total number of leaves per potÞ  100
 Z. Kheirandish, B. Harighi / Biological Control 86 (2015) 14–19 17

2.5. Statistical analysis Table 2

In vitro inhibition of growth of R. solanacearum by bacterial antagonists.

Greenhouse experiments were carried out with a factorial Strains Mean diameter of inhibition Degree of
arrangement in a completely randomized design with three repli- zone (mm)a antagonismb
cations. Data analysis was performed using the SAS software pro- Pf11 10.87b ++
gram and comparison of means was done with least significant Pf16 10.88b ++
difference (LSD) test at 5% level. Pp17 18.55a +++
Pp23 8.11c ++
Pb28 18.22a +++
En38 10.77b ++
3. Results Se40 3.99d +
R. solanacearum + sterile– 
3.1. Identification of bacterial pathogen distilled water
a
Values in the same column with the same letter(s) are not significantly different
In total, eight pathogenic isolates (Sa3, Sa35, Ta4, Kr11, Kr24, as determined by the LSD test (P = 0.01).
b
Kr48, Ba26, Pa41) were obtained from wilted potato plants in vari- Weak ‘+’ (1–5.99 mm), moderate ‘++’ (6–10.99 mm), strong ‘+++’ (11–20 mm)
inhibition effects on the growth of the pathogen.
ous locations. Isolates produced mucoid, irregular white colonies
with pink centre on TZC medium. Based on phenotypic properties
all strains were identified as R. solanacearum biovar 2.
Table 3
Furthermore, PCR using the R. solanacearum species-specific primer Siderophore, hydrogen cyanide and protease production by potential antagonistic
pairs 759/760 amplified the expected single 282-bp DNA fragment isolates.
from all the strains tested (Fig. 1). All strains were tested for
Strains Siderophore Hydrogen cyanide Protease
pathogenicity on potato and wilting was developed 14–21 days production production production
after inoculation.
Pf11 +  +
Pf16 +  +
Pp17 + + +
3.2. Identification of bacterial antagonists Pp23 +  +
Pb28   +
A total of 52 rhizobacteria were isolated from the rhizosphere of En38 +  
Se40  + 
potato plants. Isolates were preliminary identified by biochemical
and physiological characteristics and separated into six groups. + = positive,  = negative.
During preliminary screening of strains, seven with inhibiting
effects of the pathogen in vitro were selected from each group 3.4. Protease, hydrogen cyanide and siderophore production by
and further identified by partial sequencing of 16s rRNA rhizobacteria
(Table 1). The phylogenetic tree (Fig. 2) shows the position of
strains among the type strains of the species of the genera All strains, except Paenibacillus sp. Pb28 and Serratia sp. Se40
Enterobacter, Paenibacillus, Pseudomonas and Serratia. These strains were able to produce siderophore in media containing 1000 lM
were selected as potential bacterial antagonists. FeCl3. Also all strains, except Enterobacter sp. En38 and Serratia
sp. Se40 produced a clear halo zone on SKM medium, indicating
protease production. Among strains tested for Hydrogen cyanide
3.3. In vitro antagonistic activity of rhizobacteria against R. production P. putida Pp17 and Serratia sp. Se40 revealed a positive
solanacearum reaction (Table 3).

Among seven bacterial antagonists, Pseudomonas fluorescens 3.5. Greenhouse experiment

Pf11, P. fluorescens Pf16, Pseudomonas putida Pp17, Paenibacillus
sp. Pb28 and Enterobacter sp. En38 could significantly inhibit the Effects of antagonists on fresh and dry weight of shoots and
pathogen growth with mean inhibition diameter of 10.87, 10.88, roots are summarized in Table 4. There were significant differences
18.55, 18.22 and 10.77 mm, respectively. P. fluorescens Pp23 and among treatments in all the parameters evaluated. P. putida Pp17
Serratia sp. Se40 showed moderate or weak inhibition effects on showed the maximum effect on increasing both fresh and dry
the growth of the pathogen, with mean inhibition diameter of weight of shoot and root of plants. However, Serratia sp. Se40
8.11 and 3.99 mm, respectively (Table 2). was the least effective antagonist in all above parameters mea-
sured (Table 4). Surprisingly, Paenibacillus sp. Pb28 and P. putida
Pp17 were the most successful antagonists in reduction of disease
Table 1
Identification of rhizobacterial isolates by partial sequencing of 16s rRNA. by 55.56% and 51.50%, respectively compared to Serratia sp. Se40
with the least effect (Table 5).
Isolate 16s rRNA sequence % similarity GenBank
(50 ? 30 ) accession
no. 4. Discussion
Pf11 1-273, 452-1424bp 98% with Pseudomonas KP635387
fluorescens Bacterial wilt is one of the most destructive diseases of potato
Pf16 1-337, 509-1429 99% with Pseudomonas KP635388 crops worldwide. The causal agent, R. solanacearum also affect a
fluorescens
wide range of other crops such as tomato (Lycopersicon esculentum
Pp17 44-303, 349-1437 99% with Pseudomonas KP635389
putida Mill.), tobacco (Nicotiana tabacum L.), banana (Musa paradisiacal L.),
Pp23 4-258, 591-1428 98% with Pseudomonas KP635390 peanut (Arachis hypogaea L.), etc. in almost every region in the
fluorescens warm temperate, semitropical, tropical and cool climates of the
Pb28 1-251, 746-1428 99% with Paenibacillus sp. KP635386 world (Hayward, 1994). This study showed that R. solanacearum
En38 9-265, 723-1437 99% with Enterobacter sp. KP635385
Se40 6-459, 745-1437 99% with Serratia sp. KP635391
biovar 2 are responsible for bacterial wilt of potato in Kurdistan
province. This biovar had distributed worldwide and had been
18 Z. Kheirandish, B. Harighi / Biological Control 86 (2015) 14–19

Table 4
Efficacy of bacterial antagonists for potato growth promotion under greenhouse condition.

Treatment Fresh weight of shoot (g) Fresh weight of root (g) Dry weight of shoot (g) Dry weight of root (g)
a
Pf11+ R. solanacearum 34.25ab ± 1.18 4.52abc ± 0.28 4.78ab ± 0.23 0.91abc ± 0.05
Pf16+ R. solanacearum 34.83ab ± 1.02 4.47abc ± 0.21 4.88ab ± 0.20 0.90abc ± 0.10
Pp17+ R. solanacearum 36.88a ± 0.3 4.99a ± 0.16 5.17a ± 0.11 1.04ab ± 0.17
Pp23+ R. solanacearum 31.93bc ± 1.06 3.93cde ± 0.02 4.47bc ± 0.22 0.78bc ± 0.06
Pb28+ R. solanacearum 33.85ab ± 2.08 4.32bcd ± 0.17 4.69ab ± 0.31 0.88bc ± 0.04
En38+ R. solanacearum 31.21bc ± 0.18 3.74def ± 0.21 4.37bc ± 0.04 0.79bc ± 0.04
Se40+ R. solanacearum 29.24c ± 0.15 3.93ef ± 0.15 4.09 cd ± 0.07 0.79bc ± 0.05
R. solanacearum 14.92d ± 1.03 2.13 g ± 0.17 2.04e ± 0.22 0.42d ± 0.06
Controlb 29.75c ± 0.69 3.18f ± 0.03 3.74d ± 0.10 0.72c ± 0.01
a
Values in the same column with the same letter(s) are not significantly different as determined by the LSD test (P = 0.05).
b
Sterile distilled water only.

Table 5
Effect of bacterial antagonists on disease incidence and biocontrol efficacy under
greenhouse condition.
Treatment Disease incidence (%) Reduction of disease (%)
Pf11+ R. solanacearum 40.94ba ± 1.9 38.58
Pf16+ R. solanacearum 49.99c ± 1.52 25.00
Pp17+ R. solanacearum 32.33a ± 1.59 51.50
Pp23+ R. solanacearum 54.45c ± 2.85 18.31
Pb28+ R. solanacearum 29.62a ± 0.71 55.56
En38+ R. solanacearum 52.82c ± 2.71 20.76
Se40+ R. solanacearum 54.83c ± 1.86 17.74
R. solanacearum 78.40d ± 1.57 – der Weid et al. (2003), where they reported that antagonistic activ-
a ity of Paenibacillus peoriae strain NRRL BD-62 against broad spec-
Values in the same column with the same letter(s) are not significantly different
as determined by the LSD test (P = 0.05).
reported from several locations in Iran as biovar 2/race 3. (Azadvar
and Rahimian, 2006; Nouri et al., 2009). R. solanacearum can sur-
vive in soil for long periods of time. Due to its wide host range
and heterogenous nature of pathogen disease control of this patho-
gen is difficult. Then biological control by using rhizobacteria is an
efficient method for disease management. During potato plant
growth, rhizobacteria quickly colonize the underground part of
the plants as well as the rhizosphere. More than sixty bacterial
genera have been identified in different potato growing rhizo-
sphere. Among them, the species in the genus Pseudomonas such
as P. fluorescens and P. putida also to lesser extent, strains belongs
to genera Paenibacillus, Serratia and Enterobacter have high interest
as they play a role in plant protection (Diallo et al., 2011).
Furthermore other bacteria belongs to genera Bacillus had biocon-
trol efficacy on bacterial wilt of tomato plants under greenhouse
conditions (Chen et al., 2013; Tan et al., 2013). In this study several
bacteria were isolated from the rhizosphere of potato plants.
Among them, seven selected strains were further studied in vitro
and in vivo for their activity against the pathogen showing different
degree of antagonism. Among the strains tested, P. putida Pp17,
Paenibacillus sp. Pb28, P. fluorescens Pf11 and P. fluorescens Pf16
had the highest growth inhibition zone of the pathogen. Our
results of the in vitro antibiosis suggest that, inhibition effects of
these antagonists might be due to siderophore, hydrogen cyanide
and or protease production. P. putida Pp17 with the highest inhibi-
tory activity was the only strain be able to produce siderophore,
Hydrogen cyanide and protease in vitro experiments confirming
this hypothesis. There was a significant difference in in vitro inhibi-
tion of pathogen by the two P. putida Pp17 and P. fluorescens Pp23
strains. This difference might be due to inability of Pp23 strain to
produce of hydrogen cyanide. The in vitro antagonistic activity of
P. fluorescens and P. putida strains against R. solanacearum has been
previously reported (Aliye et al., 2008; Kurabachew and Wydra,
2013; Ran et al., 2005). Paenibacillus sp. Pb28 showed high inhibi-
tory activity against pathogen next to P. putida Pp17. Other pseu-
domonas species such as P. brassicacearum strain J12 with
biocontrol activity against tomato bacterial wilt had been reported.
This strain could produce hydrogen cyanide, siderophore and pro-
tease but the main active compound against R. solanacearum was
identified as 2,4-diacetylphloroglucinol (Zhou et al., 2012). There
are several previous reports about antimicrobial activity of
Paenibacillus strains against phytopathogenic bacteria and fungi
including R. solanacearum (Algam et al., 2010; Aliye et al., 2008;
von der Weid et al., 2003). Our results of in vitro experiment sug-
gest that inhibitory activity of Paenibacillus sp. Pb28 could be due
to production of protease. This finding is in agreement with von
trum of bacteria and fungi was mediated by protease production.
Enterobacter sp. En38 and Serratia sp. Se40 strains showed moder-
ate or weak inhibition effects, respectively in in vitro assay.
Antagonistic activity of Enterobacter sp. En38 and Serratia sp.
Se40 strains against pathogen might be result of production of
siderophore or hydrogen cyanide, respectively. The in vitro antago-
nistic activity of Serratia strain was previously reported by Aliye
et al. (2008) where Serratia marcescens BS-wly had weak inhibition
effect on R. solanacearum. A positive of in vitro antagonistic activity
of Serratia sp. XY21 strain and the field biocontrol efficacies against
R. solanacearum was found by Xue et al. (2013). Weak antagonistic
activity of S. marcescens SM1BA had reported by Kurabachew and
Wydra (2013) but compared to our results, analysis of in vitro pro-
duction of siderophore and hydrogen cyanide revealed positive and
negative results for SM1BA strain, respectively. In our in vivo
experiments, Paenibacillus sp. Pb28 and P. putida Pp17 significantly
reduced disease incidence by 55.56% and 51.50%, respectively.
Moreover, both strains also enhanced the plant biomass, compared
to the control. Surprisingly, P. fluorescens Pf11 and P. fluorescens
Pf16 with lower inhibition effects in vitro experiments and also
reduction of disease showed high biomass production compared
to the control. The effects of both P. fluorescens strains on enhanc-
ing biomass production, in spite of their ineffectiveness in reducing
disease might be attributed to their production of plant growth
stimulating substances more than direct inhibitory effects on
pathogen. Kloepper et al. (1980), Aspiras and Cruz (1986), Henok
et al. (2007) and Lemessa and Zeller (2007) reported that strains
of fluorescent pseudomonads significantly increased the plant bio-
mass. This could be results of induction of systemic resistance, pro-
duction of siderophore, and competition advantage to root
colonization by antagonists. Among the seven antagonistic bacteria
tested in greenhouse experiment, the highest degree of reduction
of disease was recorded by isolate Paenibacillus sp. Pb28. Also plant
biomass (fresh and dry weight of shoot and root) was increased by
this strain, compared to control. The in vivo antagonistic activity of
Paenibacillus strains and increasing effects on plant biomass under
greenhouse conditions was reported by Aliye et al. (2008). The
same results had been reported on tomato plants (Algam et al.,
2010). In conclusion, results presented in this study showed that
 Z. Kheirandish, B. Harighi / Biological Control 86 (2015) 14–19
P. putida Pp17, Paenibacillus sp. Pb28, P. fluorescens Pf11 and P. fluo-
rescens Pf16 strains with high in vitro and in vivo antagonistic activ-
ity against R. solanacearum, and also desirable plant growth
promotion effects have the potential to used as biocontrol agents.
Although, more experiments is needed to verify their effectiveness
under different field conditions or as part of disease management.
Acknowledgment
This research was supported by University of Kurdistan.
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