IOSR Journal of Agriculture and Veterinary Science (IOSR-JAVS)

e-ISSN: 2319-2380, p-ISSN: 2319-2372. Volume 9, Issue 1 Ver. I (Jan. 2016), PP 48-58
www.iosrjournals.org

Effect of bacterial isolates and phosphite compounds on disease

incidence of early blight (Alternaria solani) and improve
productivity of some potato cultivars
Agha1 M. K. M., A. I. S. Ahmed1 and S. S. Gomaa2
1
Plant Pathology Unit, Plant Protection Dept., Desert Research Center, Cairo, Egypt
2
Plant Production Dept., Desert Research Center, Cairo, Egypt

Abstract: Early blight disease caused by Alternaria solani is considered a serious challenge for potato
production. Four bacterial strains as biocontrol agents (Brevibacillus brevis, Pseudomonas
putida,Pseudomonas aerogenosa, Bacillus subtilis ) and potassium phosphite (kPhi) , copper (Cu) as well as
difenoconazole (Score) were tested to study their effects against potato early blight disease under greenhouse
and field conditions. Also the bacterial strains were tested in vitro to study their antifungal activity on growth of
A. solani. Both strains significantly reduced the fungal growth. Higher inhibition was obtained with Bacillus
subtilis while the lowest inhibition was with Brevibacillus brevis. Under greenhouse conditions, the highest
reduction was achieved with the fungicide (active ingredient difenoconazole) where disease severity reduced by
25% to 37.8% and disease incidence reduced by 36.3% to 42.9% compared to control. Bacillus subtilis (Bsu)
was the best biological agent with all potato varieties. Under field conditions, the highest reduction was
obtained with treatment by fungicide and potassium phosphite followed by Bacillus subtitles. Potato growth,
yield and its component significantly increased by fungicide and potassium phosphite compared to other
treatments. Also, Picasso cv. gave the highest shoot fresh weight and yield followed by Burren compared with
other cultivars. Potato plant fresh weight (g), average tuber weight (g) and potato yield (kg/plot) were highly
negative correlated with early blight disease incidence percent and disease severity in both seasons.
Keywords: Early blight, biological control, Alternaria solani, potassium phosphite, integrated control, foliar
spray, potato cultivars.

I. Introduction
Total world potato production is estimated at 364.8 million tonnes in 2012 (FAOSTAT, 2014). In
Egypt, potato crop has an important position among all vegetable crops, where about 20% of total area devoted
for vegetable production is cultivated with potato and the production of potato were 4.8 million tonnes in 2013
from an area of 178,000 hectares (FAOSTAT, 2014). This crop is economically important to Egypt and any
disturbance in its production affects severely it’s local and export impact. (Kabeil et al., 2008).
Early blight disease caused by Alternaria solani is considered a serious challenge for potato growing
(Van et al., 2001). In recent years, increase in disease severity on potato foliage has been reported in various
potato growing areas (Vloutoglou and Kalogerakis, 2000; Kapsa, 2003; Simões et al., 2003; Waals et al.,
2004; Pasche et al., 2005). Primary damage by early blight is attributed to premature defoliation of the potato
plants, resulting in tuber yield reduction. The pathogen can also attack potato tubers and produce shallow, dry,
corky rot (Folsom and Bonde, 1925; O’Brien and Rich, 1976). Also, early blight caused by Alternaria solani
is a major foliar disease attacking potato plants in most potato growing regions of the world. (Waals et al.,
2003;El-Gamal et al., 2007). So, using fungicidal chemical treatments to control potatoes early blight disease
mainly had been using in most regions of the world (Kapsa, 2004; Pasche et al., 2005; Mantecon, 2007).
The development of fungal resistance and problems of environmental pollution have accompanied
excessive reliance on fungicides. These problems can be avoided or minimized by using the natural material
products such as medicinal plant extracts (Chaudhary et al., 2003; Abd-El-Khair and Haggag, 2007), alkali
metal salts (Abd-El-Kareem, 2007; Labato et al., 2008), beneficial microorganisms (Wickramaarachchi,
2005; Terry et al., 2007) and cultivars resistance (Demir and Levent, 2002; Rodriguez et al., 2006; Kapsa,
2004).
Biological control agents who include effective bacterial species have been interesting as alternatives to
chemical agents (Fravel, 2005). Many species of Bacillus are known to suppress several pathogens belonging to
the genera (Cook and Baker, 1983; McKnight, 1993; Fiddaman and Rossall, 1994). On tomato plants,
Srinivasa et al., (2013) studied the effect of Pseudomonas isolates antagonistic activity against Alternaria
solani, to determine their capacity to inhibit fungal infection. And they found that, out of twelve Pseudomonas
isolates, five isolates viz., S4B7P (27.74), S1B8P (27.33), S2B10P (25.47), S3B3PF (23.07) and S1B1P (19.69)
which showed higher mean inhibition percentage on all the isolates of Alternaria solani. Also

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 Effect of bacterial isolates and phosphite compounds on disease incidence of early blight ....

Wickramaarachchi (2005) studied the effect of twenty-six exotic (including Pseudomonas fluorescens,
Azospirillum sp., P. aureofasciens [P. chlororaphis], Pseudomonas sp. and Actinomycetes [Actinomycetales])
and native rhizobacterial isolates for antagonistic activity against A. solani, and he found that, all isolates were
antagonistic to Alternaria solani.
In recent years, many scientists interested to study the relationship between the role of fertilizers either
as plant stimulator to resist or suppressor of the plant pathogens. The use of foliar fertilizers as SIR agents in
conjunction with the use of resistant varieties, improves overall control of certain diseases (Keuveni and
Reuveni, 1997). Although P fertilizer improves plant health in the majority of cases (Perrenoud, 1990). In
many cases, economic control requires periodic applications of fungicides. However, optimal fertilization can be
an important and often underappreciated component of an integrated program for disease management.
Potassium or sodium bicarbonate and Nerol showed great inhibitory effect in linear growth of
Alternaria solani. Complete inhibition was obtained with potassium or sodium bicarbonate at 2% and Nerol at
0.50 %.( Abd-El-Kareem, 2007). Phosphites (Phi) are alkali metal salts of phosphorous acid, has ability to
protect potato plants against different pathogens such as Phytophthora infestans, Fusarium solaniand and
Rhizoctonia solani (Labato, et al., 2008). Also, Keuveni and Reuveni (1998) reported that phosphates and
potassium salts effectively suppressed and controlled some diseases on various crops and they mentioned to that
foliar applied phosphite appears to be effective as a nutritional supplement and a systemic induce resistant
stimulant over a broad range of conditions.
It is cleared that, the relationship between early blight disease intensity and yield loss was highly
significant (Shtienberg et al., 1994). Some potato cultivars have resistance, while other cultivars are susceptible
(Kuczynska, 1992). So the objective of this research was to evaluate the effects of some bacterial strains as
biological control agents in vitro inhibition of Alturnaria solani in laboratory assays and determine the effect of
foliar spray of some chemical, metal salts of potassium phosphite and Cupper as mineral components with most
effective bacterial isolates on early blight disease development and suppression, yield and its components of
some potato cultivars under field conditions.

II. Materials And Methods

Bacterial strains tested as a biocontrol agents:
Tested Strains of bacteria were obtained from Microbiology Unit, Soil Fertilization and Microbiolgy
Dept. Desert Research Center, Egypt. These isolates were certified as highly effective biological control agents
against other pathogens and as biofeltlizers as mentioned in previous publication (Omar and Ahmed, 2014).

Isolation and identification of Alternaria solani:

To inoculums preparation for leaf bioassay, Alternaria solani isolates were obtained from infected
potato leaves from different growing locations (Ismailia Governorate, North Sinai governorate and Qalyubia
governorate), five isolates of A. solani were evaluated and used in this study (Table 1). The pathogens were
cultured on potato dextrose agar (PDA) in 9 cm diameter petri dishes for 10 days of incubation, the culture
covered the entire plate. The isolates were subcultured on PDA 4 days prior to use, plugs of A. solani were
formed using sterile cork borer around the perimeter of actively growing culture just before inoculation
according to (Meenakshi et al., 2014).

Table 1: Origin and year of isolation of Alternaria solani isolates

Code of isolate Location Year of isolation
As1 Al Qantara Shark 2014
As2 Al Qantara Shark 2014
As3 Baloza North Sinai 2015
As4 Baloza North Sinai 2015
As5 Qalyubia Governorate 2015

Evaluate of control agents effect on linear growth of A. solani in vitro:

Four bacterial strains as biocontrol agents (Brevibacillus brevis, Pseudomonas putida, Pseudomonas
aerogenosa, Bacillus subtilis ) were tested to study their inhibitory effects on growth of A. solani in vitro using
dual culture technique (Coskuntuna and Ozer, 2008; Alabouvette et al., 1993). Each of bacterial biocontrol
agents was streaked in center of sterile Petri dish on potato dextrose agar (PDA). One disc (0.5 cm in diameter)
of 7 days – old culture of each pathogenic fungus was separately placed on each side of the same petri dish at 10
mm distance. Petri dishes containing of 15 ml PDA medium inoculated with fungal cultures and free of bacteria
were used separately as control. All plates were incubated at 28 ±2ºC until the growth of each pathogenic fungus
in the control treatment reached to the edge of Petri dish. The inhibition growth rate was calculated by following
equation: Inhibition growth (%) = ( R – r) / r X 100 Where (R) is the diameter of mycelial growth fungi on the
control cultures and (r) is the growth diameter of fungal mycelia on the plate treated with bacterial isolates.
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 Effect of bacterial isolates and phosphite compounds on disease incidence of early blight ....

Preparation of fungal suspension:

The fungus A. solani was grown on PDA medium at 25±2°C until an abundant heavy growth of conidia
was evident. Conidia were harvested by scraping the surface of the colonies with a spatula and transferred to
sterilized distilled water and filtered through nylon mesh to prepare the suspension of pathogen.

Preparation of Biocontrol agents:

Bacterial strains were grown on nutrient broth medium and incubated for 48h at room temperature in a
rotary shaker (100 round/min). The bacterial culture was centrifuged (10000 rpm for 10 min) and the
supernatant was discarded. The cell pellet was suspended in sterile 0.85% NaCl and centrifuged again under the
same conditions. The supernatant was discarded and washed bacterial cells were re-suspended in sterile distilled
water. The cell concentrations was adjusted to 108CFU/ml and used for field and greenhouse experiments.

Chemical compounds:
- Potafore ―a commercial compound containing 42% potassium and 38 % phosphorus as potassium phosphites
(kPhi)‖ and sprayed by 2 ml/1L water.
- Oxy plus ―a copper compound contain copper oxychloride by 47.89%‖ and sprayed by 2.5 gm/1L water.
- Score ―a fungicide contained difenoconazole and used by 0.5 ml / 1L water.

Greenhouse experiment:
The experiment was carried out in the greenhouse of plant protection department, Desert Research
Center, Egypt. Potato tubers cvs. Burren, Diamond, Picasso and Spunta were grown in plastic pots (50 cm
diameter) containing sandy loam soil at 22–25°C and RH 75–80%. Five plants/pot and three pots for each
treatment were used. Plant inoculation was carried out with 30 ml of fungus suspension when plants had 4–5
leaves. After inoculation, pots kept covered under plastic sheet for two days and treated with 90 ml from
bacterial strain with concentration of bacterial cells (108CFU/ml.) for each isolate. while the concentration of
other compounds were (2 ml/L, 2.5 gm/L and 0.5 ml/L) for Potafore, Oxy plus and Score respectively.
Inoculated potato plants treated with water were used as control. Disease incidence and severity were rated
based on percentage of damaged potato leaf area and affected number of plants.

Field Experiments (Experimental site, cultivars and cultivation)

Field experiment was conducted under conditions of Baloza experimental station, desert research
center, North Sinai governorate, Egypt, during summer growing seasons of 2014 and 2015 to evaluate the
efficacy of bacterial strains of Brevibacillus brevis (Bbr), Pseudomonas putida (Ppu), Pseudomonas aerogenosa
(Pae), Bacillus subtilis (Bsu) as biological control agents and Potassium phosphite (kPhi) , Copper (Cu) as well
as difenoconazole (Score) on incidence and severity of early blight disease on four potato commercial cultivars
( Burren, Diamond, Picasso and Spunta) under natural conditions. All treatments were used as a foliar spray 30
days after emergence (4 –5 true leaf stage), four times with 15 days interval, the concentration of treatments
were (108 CFU/ml ; 2 ml/L, 2.5 gm/L and 0.5 ml / L) for bacterial isolates, Potafore, Oxy plus and Score
respectively, the same amount of water was sprayed to the control treatment to avoid interferences with different
moisture levels. This experiment was conducted in split plot design with three replicates where potato cultivars
assessed in main plot while foliar spry treatments were assessed in sub plot. Exported potato seed tubers of four
cultivars planted at 15th January in rows 1 m width, 30 cm apart under drip irrigation system and the plot area
was 10.5 m2. In both seasons, all cultural practices (irrigation, fertilization, weeding, and pest control) were
performed according to the recommendations of the Egyptian Ministry of Agriculture and Land Reclamation.

Plant traits and disease assessments:

Vegetative growth:
Random samples of five plants of each experimental plot were taken at 70 days after planting for
vegetative growth data. Plant height, leaves number/plant, leaves chlorophyll content and plant fresh weight
were recorded.

Yield and its components:

120 days after planting all experimental plots were harvested then tubers were counted, and weighed to
record of total yield, tuber number and average tuber weight.
At the end of each growing season, the average harvested yield was calculated as kg/plot.

Disease incidence and severity:

These reading were recorded with the appearance of natural disease symptoms on control plants (60
days after planting). Early blight incidence was recorded according to the number of infected plants showed

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 Effect of bacterial isolates and phosphite compounds on disease incidence of early blight ....

disease symptoms in relation to the whole number of potato plants. The average of records of the surveyed
replicates for each particular treatment was calculated. Disease severity and incidence were estimated as
follows; 0 = no leaf lesion; 1= lesions on < 25% of leaf area; 2 = lesion on 26–50% of leaf area; 3 = lesion on
51–75% of leaf area and 4 = lesions on 76 up to 100% of leaf area according to the scale from 0 to 4 suggested
by Cohen et al., (1991) then the traits calculated using the following formula:

∑ (n×c)

D.S = ________
N

Where; D.S. = intensity of attack, n= number of infected plants per category, c = category number and N = total
examined plants.
Statistical analysis:
Data were subjected to statistical analysis by M-STAT C (Russel, 1991). The differences among means were
performed using least significant difference (LSD) at 5% level.

III. Results And Discussion

Effect of biocontrol agents on linear growth of Alternaria solani in vitro:

Four bacterial strains were used to study their inhibitory effect on growth of Alternaria solani in vitro.
Results indicated that both treatments reduced the linear growth of Alternaria solani. Higher inhibition was
obtained with Bacillus subtilis in all cases of tested fungal isolates while the lowest inhibition rate with
Brevibacillus brevis. The most affected pathogenic isolates was As2 followed by As1 in case of Bacillus
subtilis which reduced the linear growth by 65.67% and 62.5 % respectively. On the other hand, the As1 was
most resistant under stress of Brevibacillus brevis where recorded 25.5% inhibition rate (Fig 1). These results
are in agreement with (Edwards et al., 1994; Fravel, 2005; Sallam and Abo-Elyousr, 2012; Stepanović et al.,
2015 ). As preliminary results of screening for antagonism in petri plates assay, all tested bacterial isolates were
selected to evaluate their antifungal effects in further experiments in compared to the effect of fungicide (with
active ingredient Difenoconazole, potassium phosphite (kphi) and Cupper (cu).

Fig. 1: Linear growth of Alternaria solani as affected with bacterial isolates in vitro

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 Effect of bacterial isolates and phosphite compounds on disease incidence of early blight ....

Influence of foliar spray treatments on early blight diseases of potato under greenhouse conditions:

To determine the activity of biocontrol agents (Brevibacillus brevis, Pseudomonas putida,

Pseudomonas aerogenosa, Bacillus subtilis), Potassium phosphite (kphi), Copper (Cu) as well as
difenoconazole on early blight disease under greenhouse conditions, the concentrations were applied on potato
leaves after inoculation of the pathogens. The results in Tables (2 and 3) showed significant reductions in
disease severity and incidence. The most effective treatments were in case of fungicide with all potato cultivars
where the percentage of disease severity reduced by 37.8%, 32.5%, 32.5%, 25% with cvs. Burren, Diamond,
Picasso and Spunta respectively as a reduction rates between control and treatments followed by Bacillus
subtilis (Bsu) which gave the best biological agent effect with all potato cultivars except Diamond, where the
reduction percentage of severity by Bsu were 27%, 32.5% and 25% in Burren, Picasso and Spunta, respectively
compared to control. Also the disease incidence induction rates were compatible with disease severity results
where the incidence has been reduced by 36.3%, 38.5%, 42.9% for cvs. Burren, Diamond, Picasso and Spunta
respectively, when treated with pesticide. Also (Bsu) gave reduction rates of 42.87%, 30.80% with Picasso and
Spunta, respectively. Concerning of biological agents, no significant differences between Brevibacillus brevis
and Pseudomonas putida. The reduction rate was between 17% with Picasso to 7.5% with Diamond and Spunta.
As expected, the chemical treatment using fungicide were the best to reduction of disease under greenhouse
conditions followed by the bacterial isolates and phosphite compounds, so these results led to use all tested
treatments under greenhouse to combat early blight disease and reduce the use of fungicides, these results
agreed with (Westermann, 1993; Shtienberg et al., 1996; Christ, 1990; Hofman et al., 2003; William et al.,
2007).

Table (2): Evaluation of biological and chemical control agents against Alternaria solani under
greenhouse conditions.
Burren Diamond Picasso Spunta X¯
Treatments DI DS DI DS DI DS DI DS DI DS
Bbr 66.7 3.3 73.3 3.7 73.3 3.7 73.3 3.7 71.7 3.6
Ppu 66.7 3.3 73.3 3.7 66.7 3.3 73.3 3.7 70.0 3.5
Pae 60.0 3.0 80.0 4.0 73.3 3.7 80.0 4.0 73.3 3.7
Bsu 53.3 2.7 66.7 3.3 53.3 2.7 60.0 3.0 58.3 2.9
(Kphi) 66.7 3.3 53.3 2.7 60.0 3.0 66.7 3.7 61.7 3.2
Cu 60.0 3.0 60.0 3.0 60.0 3.0 66.7 3.3 61.7 3.1
Score 46.7 2.3 53.3 2.7 53.3 2.7 60.0 3.0 53.3 2.7
Control 73.3 3.7 86.7 4.0 93.3 4.0 86.7 4.0 85.0 3.9
X¯ 61.7 3.1 68.3 3.4 66.7 3.3 70.8 3.5
LSD at 5%
Characters Cultivars Foliar spray Interaction
Dis. Inci. 7.4 8.0 16.0
Dis. Sev. 0.3 0.4 0.7

Potato cultivars response towards Alternaria solani infection and foliar spray under field conditions:
The efficacy of four bacterial isolates and chemical components on disease incidence and severity
under field conditions are shown in Tables (3 and 4). Results revealed that all cultivars have deferent susceptible
to early blight disease incidence and severity recorded during two seasons. Burren cultivar exhibit less
susceptibility to early blight according to disease incidence and severity compared with other cultivars.
On the other hand all foliar spray treatments gave significant results with Burren cultivar in both
seasons. Picasso cultivar seems to be moderate susceptible in both seasons (54.3% and 44.8%) (3.3 and 3.0) for
disease incidence and severity in first and second seasons respectively. Cultivars Diamond and Spunta showed
more susceptibility to early blight in both seasons. Diamond seems to be more susceptible than Spunta in first
season (63.8% and 62.9%) (4 and 3.7) for disease incidence and severity with control treatment. In the same
trend, Spunta showed more susceptible for disease incidence in second season than Diamond especially with
control where recorded (52.4% and 48.6%) for both cultivars.
Concerning foliar spray, all foliar spray treatments gave significantly decreased disease incidence and
disease severity compared with control treatment in both seasons.
Bacterial isolates Brevibacillus brevis (Bbr) Pseudomonas putida (Ppu), Pseudomonas aerogenosa
(Pae) and Bacillus subtilis (Bsu) gave significant results cambered to control. While fungicide (Score),
Potassium phosphite (kphi) and copper (cu) gave the highest significant reduction in disease incidence and
severity compared with bacterial isolates and control treatments. (Bsu) was more effective in decreasing the
disease incidence and severity than other isolates with all cultivars through two seasons where, recorded (31.7
and 26.7) and (2.7 and 1.8) of disease incidence and severity in first and second seasons, respectively. Treated

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potato plants with Potassium phosphite (kphi) and Copper (Cu) gave superior results for disease incidence and
severity compared with bacterial isolates and control (Tables 3 and 4). The highly decreasing in disease
incidence and severity obtained with Score treatment compared with other treatments. These results are
agreement with (Kapsa and Osowski, 2003; Kapsa, 2004; Pasche et al., 2005; Mantecon, 2007).

Table (3): Effect of cultivars and foliar spray treatments and their interactions on potato disease
incidence during two successive growing seasons (2014 and 2015).
Burren Diamond Picasso Spunta X¯
Treatments 1St S* 2nd S** 1St S 2nd S 1St S 2nd S 1St S 2nd S 1St S 2nd S
Bbr 35.2 38.1 44.8 40.0 42.9 41.0 46.7 42.9 42.4 40.5
Ppu 34.3 35.2 42.9 40.0 41.0 39.0 43.8 44.8 40.5 39.8
Pae 31.4 32.4 41.0 41.9 42.9 41.9 42.9 41.9 39.5 39.5
Bsu 21.0 21.0 35.2 31.4 32.4 23.8 38.1 30.5 31.7 26.7
(Kphi) 19.0 21.0 32.4 30.5 21.0 21.0 34.3 28.6 26.7 25.2
Cu 21.9 22.9 28.6 26.7 26.7 21.0 29.5 31.4 26.7 25.5
Score 10.5 5.7 20.0 18.1 16.2 11.4 19.0 15.2 16.4 12.6
Control 43.8 41.9 63.8 48.6 54.3 44.8 62.9 52.4 56.2 46.9
X¯ 27.1 27.3 38.6 34.6 34.6 30.5 39.6 36.0
LSD at 5%
Cultivars Foliar spray Interaction
1St S 1.8 1.9 3.8
2nd S 2.2 3.0 5.9
*, ** Means the first (1st S) and second (2nd S) growing seasons

Table (4): Effect of cultivars and foliar spray treatments and their interactions on potato disease severity
during two successive growing seasons (2014 and 2015).
Burren Diamond Picasso Spunta X¯
Treatments 1St S* 2nd S** 1St S 2nd S 1St S 2nd S 1St S 2nd S 1St S 2nd S
Bbr 2.3 1.7 3.7 1.7 2.7 2.3 3.3 2.7 3.0 2.1
Ppu 2.0 2.0 3.7 2.7 2.3 2.3 2.7 3.0 2.7 2.5
Pae 2.3 2.3 4.0 3.0 3.0 2.7 3.3 2.7 3.2 2.7
Bsu 1.7 1.3 3.3 1.7 2.3 1.7 3.3 2.3 2.7 1.8
(Kphi) 1.7 1.7 3.0 2.3 2.7 1.7 3.7 2.3 2.8 2.0
Cu 1.7 1.3 3.0 2.0 2.3 1.3 3.0 2.0 2.5 1.7
Score 1.3 1.0 2.3 1.7 1.7 1.0 2.3 1.3 1.9 1.3
Control 2.7 2.7 4.0 3.3 3.3 3.0 3.7 3.0 3.4 3.0
X¯ 2.0 1.8 3.4 2.3 2.5 2.0 3.2 2.4
LSD at 5%
Cultivars Foliar spray Interaction
1St S 0.6 0.4 0.9
2nd S 0.2 0.4 0.9
*, ** Means the first (1st S) and second (2nd S) growing seasons. Early Blight Disease Severity Recorded
according to the Scale 0-4;

Efficacy evaluating of foliar spray application of bacterial biocontrol and chemical control agents on
early blight disease under Field condition:
To evaluating the efficacy of treatments on tested potato cultivars vegetative growth, data presented in
(Table 5) showed that potato shoot length, shoot fresh weight, leaf number and leaf chlorophyll content
significantly affected by potato cultivars and foliar spray treatments in both seasons except the effect of cultivars
in the second season. Moreover, interaction effects were not significant except their effect on shoot fresh weight
in both seasons. Picasso cv produced the highest shoot length and highest shoot weight in both seasons
compared with other cultivars, followed by Diamond for shoot length, Burren and Spunta for shoot fresh weight
in both seasons. The lowest shoot length was obtained by cvs Burren and Spunta, while cv Diamond produced
the lowest shoot fresh weight in both seasons. Burren and Diamond cultivars gave the highest leaf number
compared with cv Spunta in both seasons, while the highest leaf chlorophyll content was obtained by cv Burren
compared with other cultivars in both seasons.
Concerning foliar spray effects, data in (Table 5) showed that Score treatment (fungicide) significantly
increased shoot length, shoot fresh weight and leaf number compared with other treatments, followed by
potassium phosphite (kphi) then )Bsu( bacterial strain. While control treatment produced the lowest values
followed by other bacterial strains and Cupper treatments in both seasons. Highest leaf chlorophyll content was
obtained with potassium phosphite, Score and Cupper treatments compared with control and bacterial strains
treatments in both seasons. Picasso cultivar treated with Score gave the highest shoot fresh weight, while other
cultivars with control treatment gave the lowest value in both seasons. Similar results were obtained by
(Kuczynska, 1992; Keuveni and Reuvcni, l997; Keuveni and Reuvcni, 1998).
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 Effect of bacterial isolates and phosphite compounds on disease incidence of early blight ....

Table (5) Effect of cultivars, foliar spray treatments and their interactions on potato shoot length,
fresh weight, leaves number and leaves chlorophyll content in two seasons.
characters Shoot length (cm) Shoot fresh weight (g) Leaves number chlorophyll (SPAD)
Seasons St nd St nd St nd
1 S 2 S 1 S 2 S 1 S 2 S 1St S 2nd S
Cultivars
Burren 58.08 60.38 304.31 266.40 21.35 17.91 46.96 52.28
Diamond 64.71 67.29 243.15 243.15 20.98 18.53 43.68 49.75
Picasso 70.84 73.87 380.62 380.62 20.10 17.40 44.24 50.10
Spunta 57.90 60.54 301.88 301.88 18.89 16.03 44.91 50.31
LSD at 0.05 2.08 1.60 20.31 17.22 1.47 0.93 1.68 NS
Foliar spray
Bbr 61.24 64.26 291.12 281.56 18.56 15.57 44.17 50.15
Ppu 60.57 62.37 286.73 279.18 19.08 16.09 43.66 49.56
Pae 61.48 63.61 292.89 283.32 19.75 16.68 43.47 49.12
Bsu 61.98 65.42 309.18 303.68 20.55 17.64 44.21 50.69
(KPhi) 67.31 69.86 360.58 352.59 21.88 19.64 47.49 53.14
Cu 60.33 62.21 286.08 276.43 20.33 17.34 46.30 50.86
Score 71.07 75.29 401.65 383.66 23.90 20.41 46.92 52.58
Control 59.08 61.13 231.70 223.69 18.61 16.37 43.36 48.76
LSD at 0.05 2.34 2.39 13.57 14.43 1.46 1.23 1.96 1.67
Interaction
Bbr 55.74 58.88 307.02 268.77 20.21 16.89 46.18 52.25
Ppu 55.09 56.89 302.00 271.80 20.58 17.26 45.80 51.87
Pae 56.34 58.16 309.27 270.97 20.86 17.54 44.67 50.07
Burren

Bsu 56.65 60.45 305.13 283.17 21.59 17.61 45.95 52.35

(KPhi) 60.54 63.67 321.73 289.77 22.30 19.65 50.23 55.63
Cu 59.70 59.50 317.43 278.80 20.58 17.59 48.21 51.28
Score 65.05 68.18 365.23 293.27 25.61 19.62 48.68 53.42
Control 55.50 57.30 206.70 194.67 19.08 17.09 45.97 51.37
Bbr 65.78 68.58 220.90 220.90 20.29 16.97 43.25 49.32
Ppu 64.29 66.09 201.83 201.83 19.97 17.31 42.19 48.26
Diamond

Pae 63.60 65.40 209.97 209.97 20.12 17.80 43.13 48.53

Bsu 65.75 69.78 236.60 236.60 21.81 18.49 43.92 50.99
(KPhi) 68.14 71.61 316.03 316.03 22.74 21.09 45.07 52.13
Cu 61.35 63.15 214.93 214.93 19.80 17.48 44.23 48.63
Score 69.72 72.18 347.00 347.00 22.76 21.44 45.99 53.06
Control 59.02 61.48 197.97 197.97 20.32 17.67 41.68 47.08
Bbr 68.95 71.65 351.33 351.33 17.34 14.69 43.55 49.95
Ppu 68.60 70.40 368.87 368.87 18.93 15.61 43.57 49.63
Pae 69.92 71.72 361.20 361.20 20.24 16.92 42.26 48.66
Picasso

Bsu 69.18 70.98 381.03 381.03 17.98 16.99 43.13 50.19

(KPhi) 77.27 79.07 422.43 422.43 21.60 19.61 47.30 52.03
Cu 64.22 68.36 335.77 335.77 21.29 17.97 46.27 51.67
Score 81.62 90.08 497.20 497.20 24.22 20.90 45.24 50.64
Control 66.92 68.72 327.10 327.10 19.19 16.54 42.60 48.00
Bbr 54.48 57.95 285.23 285.23 16.40 13.75 43.69 49.09
Ppu 54.31 56.11 274.20 274.20 16.84 14.19 43.07 48.47
Pae 56.02 59.15 291.13 291.13 17.79 14.47 43.83 49.23
Spunta

Bsu 56.33 60.46 313.93 313.93 20.80 17.48 43.84 49.24

(KPhi) 63.29 65.09 382.13 382.13 20.87 18.21 47.37 52.77
Cu 56.02 57.82 276.20 276.20 19.63 16.31 46.48 51.88
Score 67.91 70.71 397.17 397.17 22.99 19.67 47.79 53.19
Control 54.87 57.01 195.03 195.03 15.83 14.18 43.21 48.61
LSD at 0.05 NS NS 27.14 28.87 NS NS NS NS

After confirming the effectiveness of treatments in reducing the disease incidence and severity of
potato late blight, the component of yield were investigated, data presented in (Table 6) showed that potato yield
per plot, tuber number, average tuber weight and tuber dry matter percent significantly affected by tested potato
cultivars and foliar spray application in both seasons. Only tuber number per plot in the second season was not

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 Effect of bacterial isolates and phosphite compounds on disease incidence of early blight ....

significant. Moreover, all interaction effects (Table 7) were not significant except total yield per plot in both
seasons and tuber number per plot in the first season. Picasso cultivar gave the highest potato yield, followed by
cv Burren compared with cvs Spunta and Diamond which gave the lowest yield in both seasons.
Picasso cultivar produced the highest tuber number per plot followed by Diamond in the first season
compared with cv Spunta which produced the lowest tuber number per plot. On the other hand, cvs Spunta,
Burren and Picasso in the first season, Spunta and Picasso in the second season produced the highest tuber
weight compared with cv Diamond which gave the lowest value in this respect. Regarding dry matter percent,
cvs Picasso and Diamond gave the highest percent compared with cvs Spunta and Burren in both seasons.
Concerning of foliar spray treatments, Score treatment significantly increased potato yield, tuber
weight and tuber dry matter percent followed by potassium phosphite then (Bsu) bacterial strain compared with
other treatments. Picasso cultivar treated with Score gave the highest potato yield compared with cv Diamond
treated with Cupper, bacterial strains and control treatments in both seasons.

Table (6) Effect of cultivars and foliar spray treatments on potato yield (kg/plot), tubers number per plot,
average tuber weight (g) and dry matter % in both of two growing seasons.
Potato yield (kg/plot) Tubers number/plot Average tuber weight (g) Dry matter %
Cultivars
1St S* 2nd S** 1St S 2nd S 1St S 2nd S 1St S 2nd S
Burren 29.88 25.92 285.1 270.5 104.90 96.39 18.82 16.87
Diamond 26.50 22.54 295.4 279.1 89.58 81.50 22.11 19.51
Picasso 34.80 30.92 333.9 288.60 104.35 108.50 22.78 19.34
Spunta 28.09 24.87 261.8 245.4 106.82 101.22 20.72 18.08
LSD at 0.05 3.57 2.75 43.1 NS 2.87 9.94 0.97 0.72
Foliar spray
Bbr 28.39 24.17 287.76 270.40 98.67 89.83 20.39 17.56
Ppu 28.18 23.63 287.32 270.58 98.53 88.49 20.66 17.87
Pae 28.44 23.89 287.74 269.31 98.94 89.05 20.59 18.07
Bsu 30.01 27.13 298.54 278.95 100.41 97.69 21.34 18.40
(KPhi) 34.78 30.82 323.81 283.23 107.76 109.32 21.94 19.61
Cu 27.68 23.55 278.56 256.89 99.39 92.13 20.94 18.15
Score 36.40 32.60 333.64 287.50 109.69 115.00 22.08 19.58
Control 24.67 22.70 254.93 250.32 97.91 93.27 20.90 18.38
LSD at 0.05 1.25 1.37 19.1 NS 4.00 8.16 0.81 0.54
*, ** Means the first (1St S) and second (2nd S) growing seasons.

Relationship between early blight disease control and yield components of tested potato cultivars:
Based on the obtained results, it may be worth to mention that potato plant fresh weight (g), average
tuber weight (g) and potato yield (kg/plot) were highly negative correlated with early blight disease incidence
percent in both seasons, fig.(1). A linear correlation coefficients (r) were (-0.599, -0.540), (-0.487, -0.570) and (-
0.593, -0.636) for plant fresh weight, average tuber weight and potato yield in the first and second seasons,
respectively. The corresponding coefficient of determination (r 2) was (0.359, 0.292), (0.337, 0.327) and (0.352,
0.404), which indicated that (35.9%, 29.2%), (23.7%, 32.7%) and (35.2%, 40.4%) of the variation in plant fresh
weight, average tuber weight and potato yield were related to early blight disease incidence percent in the first
and second seasons, respectively. On the other hand regression coefficients were (-3.38, -3.43), (-0.31, 0.71) and
(-0.25, -0.28) for plant fresh weight (g), average tuber weight (g) and potato yield (kg/plot) in the first and
second seasons, respectively. This indicated that when early blight disease incidence increased by one percent,
plant fresh weight, average tuber weight and potato yield decreased by (3.38g, 3.43 g), (0.31g, 0.71g) and (0.25
kg, 0.28 kg) in the first and second season, respectively fig (2).

Moreover, the same characters were highly negative controlled with early blight disease severity in
both seasons fig (2). Whereas, a linear correlation coefficients (r) were (-0.586, -0.449), (-0.573, -0.457) and (-
0.561, -0.523) for plant fresh weight (g), average tuber weight (g) and potato tuber yield (kg/plot) in the first and
second seasons, respectively. The corresponding coefficients determination (r 2) were (0.343, 0.201), (0.328,
0.209) and (0.315, 0.273) which indicated that (34.3, 20.1), (32.8, 20.9) and (31.5, 27.3) of the variation in plant
fresh weight, average tuber weight and potato yield were related to early blight disease severity in the first and
second seasons, respectively.

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 Effect of bacterial isolates and phosphite compounds on disease incidence of early blight ....

Table (7): The interactions between cultivars and foliar spray treatments on potato yield (kg/plot), tubers
number per plot and average tuber weight (g) of two seasons
Potato yield (kg/plot) Tubers number/plot Average tuber weight (g) Dry matter %
Characters/
seasons
1St S* 2nd S** 1St S 2nd S 1St S 2nd S 1St S 2nd S
Cultivars/
treatments
Bbr 29.33 24.78 285.80 274.51 102.34 89.86 18.22 15.58
Ppu 29.28 24.73 288.56 279.64 102.13 89.83 18.44 16.45
Pae 29.12 24.57 285.39 274.81 102.54 90.37 18.47 16.55
Burren

Bsu 29.86 27.97 287.76 275.10 103.77 102.05 19.26 17.17

(KPhi) 32.53 27.98 292.37 271.54 111.37 103.28 19.68 17.95
Cu 29.49 24.94 285.96 274.12 103.00 90.88 18.42 16.42
Score 34.78 31.57 309.70 284.00 112.46 111.90 19.52 18.30
Control 24.69 20.81 244.83 230.43 101.61 92.99 18.53 16.56
Bbr 23.50 18.95 274.59 273.38 86.68 72.42 20.33 18.38
Ppu 22.69 18.14 263.38 257.33 86.73 72.10 21.56 18.99
Pae 23.10 18.55 265.37 257.11 87.14 72.51 21.30 18.68
Diamond

Bsu 24.84 22.96 278.47 268.68 89.37 86.54 21.86 18.57

(KPhi) 33.00 28.45 343.89 295.73 95.97 96.70 23.46 20.99
Cu 23.52 19.97 270.67 258.75 87.60 79.17 22.74 19.67
Score 35.45 30.90 365.35 318.66 97.06 97.17 23.66 20.83
Control 25.92 22.37 301.23 302.81 86.08 75.34 21.97 19.97
Bbr 34.31 31.10 337.14 297.53 101.59 104.66 22.76 18.64
Ppu 35.42 30.87 351.74 316.45 101.38 99.60 22.27 18.24
Pae 34.51 29.96 340.18 301.45 101.79 100.45 22.30 19.41
Picasso

Bsu 36.06 31.51 350.25 310.28 103.02 101.78 23.52 19.65

(KPhi) 37.49 33.94 339.30 279.52 110.62 122.05 23.24 20.32
Cu 32.20 27.65 314.36 270.76 102.25 101.88 22.19 18.77
Score 37.73 34.85 333.88 270.68 113.38 131.50 23.73 20.92
Control 30.67 27.45 304.73 262.15 100.73 106.11 22.23 18.79
Bbr 26.42 21.87 253.51 236.17 104.06 92.40 20.24 17.63
Ppu 25.32 20.77 245.61 228.88 103.85 92.41 20.38 17.79
Pae 27.01 22.46 260.02 243.85 104.26 92.88 20.31 17.65
Spunta

Bsu 29.29 26.08 277.70 261.71 105.49 100.39 20.71 18.19

(KPhi) 36.11 32.90 319.67 286.14 113.09 115.25 21.37 19.16
Cu 25.52 21.64 243.23 223.92 104.72 96.57 20.41 17.76
Score 37.62 33.07 325.62 276.64 115.85 119.45 21.40 18.26
Control 17.40 20.19 168.93 205.87 103.20 98.63 20.89 18.22
LSD at 0.05 2.50 2.73 38.1 NS NS NS NS NS

The regression coefficients were (-57.29, -50.87), (-6.39, -9.99) and (-4.01, -4.06) for plant fresh
weight, average tuber weight and potato yield which indicated that when early blight disease severity increased
by one grade on scale, plant fresh weight , average tuber weight and potato tuber yield decreased by
(57.29g,50.87g), (6.39g, 9.99g) and (4.01kg, 4.06kg) in the first and second seasons, respectively fig (2). Such
decrement of potato yield /plot due mainly to average of tuber weight which in turn, decreased when plant fresh
weight especially leaves (assimilative plant organs) were significantly affected by early blight disease
incidence and severity. Similar results were found by (Folsom and Bonde, 1925; O’Brien and Rich, 1976;
Vloutoglou and Kalogerakis, 2000; Simoes et al., 2003; Waals et al., 2004; Pasche et al., 2005; El-Gamal et
al., 2007; Amitava and Devi, 2014) where they reported that primary damage by early blight is attributed to
premature defoliation of the potato plants, resulting in tuber yield reduction.

IV. Conclusion
The results of this study showed that the tested biological, chemical and fertilizing factors exhibited
different levels of antifungal activities on A. solani isolates under laboratory, greenhouse and field conditions.
The highest reduction was achieved with the Score fungicide (active ingredient Difenoconazole), and Bacillus
subtilis was the most effective bacterial isolates under vitro and vivo conditions. There is a relation between
yield production and rate of disease infection either severity or incidence, and the treatments were effective in
reduce of infection rate and increase of potato yield, so we conclude that, using of Bacillus subtilis as biocontrol
agents and with integrated management program with potassium phosphite or fungicide may be useful in
controlling potato early blight disease under field conditions.

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 Effect of bacterial isolates and phosphite compounds on disease incidence of early blight ....

Fig.(2) Regression lines, coefficients of determination (r2) and regression coefficients (b) for plant fresh
weight (g/plant), total yield(kg/plot) and average tuber weight (g) of potato vs. early blight (A)
disease incidence % and (B) disease severity for 2014 and 2015 seasons.

Acknowledgements:
The authors would like to thank Associate Prof. Amal M. Omar, Microbiology Unit, Soil Fertilization
and Microbiology Dept. Desert Research Center, Egypt, for providing the bacterial isolates which used in this
research work.

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