Journal of Phytopathology and Pest Management 8(1): 15-28, 2021

pISSN: 2356-8577 eISSN: 2356-6507

Journal homepage: http://ppmj.net/

∗

Corresponding author:

Ramadan R. A. Hussein,

E-mail: ramadanhussein.5419@azhar.edu.eg

15

Effect of some biofertilizers and biofungicides

applications on control onion root-rot disease

Ramadan R. A. Hussein

*

, Mohamed M. El-Sheikh Aly, Abd-Elal A. Mohamed

Agricultural Botany Department, Faculty of Agriculture, Al-Azhar University, 71524 Assiut, Egypt

Abstract

Keywords: onion, Allium cepa, root rot, biofertilizers, biofungicides.

16

1. Introduction

Onion (

Allium cepa

L.) is a species of the

alliaceae family, the largest commercial

crop in the world and used for food and

medicine since ancient times (Marrelli et

al. 2019; Yang et al. 2019; Hossain et al.

2017; Khan et al. 2017). China, India,

USA, Turkey, Japan, Spain, Brazil,

Poland, and Egypt are the largest

countries of the world producing onion

(FAO, 2018). Egypt’s production of

onion reached 2.4 million tons (FAO,

2017). In Egypt, Onion to the second

important cash crop after rice. The

consumption is attributed to several

factors, which mainly heavy promotion

that links flavor and health and the

popularity of onion-rich ethnic foods. The

litle production of onion is due to effects

of fertilizers, growing unsuitable varieties

and different root-rots fungi diseases

under the agro climatic conditions of an

area. Onion plants are infected with a

wide spectrum of fungal diseases as they

are 23 diseases as surveyed (Conn et al.,

2012). In Egypt, onion plants are most

infected with fungal diseases than any

other diseases due to climatic conditions

which are suitable to infect the plant. In

Egypt, Onions suffered from the fungal

root-rots the most prevalent and

dangerous root disease all over the

country and world wide

.

However, yield

losses reached 75-80% in case of soil

born fungi (Tjamos et al., 2010). This

work aimed to study the effect of some

bio-fertilizers and biofungicides on

control onion root rot disease.

2. Materials and methods

This work was carried out in the

Research Laboratory and Farm of Faculty

of Agriculture, Al-Azhar University

(Assiut Branch), Assiut, Egypt.

2.1 Isolation and identification of the

causal pathogens

Survey for root-rot diseases were

conducted during season 2016/2017 at

different localities in Menofeia, Assiut,

Sohag and Luxor on onion plants.

Samples of diseased plants were taken

into the laboratory to isolate the causal

pathogens. The infected roots were

excised and carefully washed with tap

water to remove any adhesive soil. Small

segments of the infected roots were

superficially sterilized in 70% ethyl

alcohol for 2 minutes. Then, the

fragments were left to dry on sterilized

filter papers then placed on PDA plates

and incubated at 27 0c for seven days.

The isolated fungi were purified using

the single spore or the hyphae tip

technique (Dhingra and Sinclair, 1985).

The purified fungi were identified

according to fungal morphological and

microscopical characteristics as

described by Barnett and Hunter (1986),

Booth (1977)

and confirmed by

Agricultural Botany Department, Faculty

of Agriculture, Al-Azhar University

(Assiut branch), Egypt. The obtained

isolates were maintained on PDA slants

and kept in refrigerator at 5°C for further

studies. The frequency of the isolated

fungi was calculated separately for each

of the collected samples. Stock cultures

were routinely sub-cultured on fresh

slant every month.

2.2 Pathogenicity tests

Pathogenic capability of 40 isolates was

carried out on onion transplants (Giza 6

Mohassan cv.) under greenhouse

conditions, Faculty of Agriculture, Al-

Azhar University, Assiut Egypt during

17

2016/2017 growing season. Pots (30 cm

diameter) were sterilized by immersing in

5 % formalin for 15 minutes, and then

left to air dry for 10 days. A mixture of

clay soil and sand (1:1 w/w) was also

sterilized with the same solution and

covered with polyethylene sheet for 15

days, then left uncovered to remove the

residual of formalin for 10 days. The pots

were filled with the sterilized soil (5

kg/pot).

2.3 Inoculum preparation

The fungal inoculum was grown in 250

ml plastic jars containing the following

substrate per jar (75 g grain barley, 25 g

coarse sand and 25 ml tap water to cover

the mixture in jars). The jars were

autoclaved at 121°C for 30 minutes, left

to cool, then inoculated with the tested

fungi and incubated at 25°C for 15 days

to obtain sufficient growth of each

fungus. Then, sterilized plastic pots in

5% formalin solution (30 cm in diameter)

were filled with sterilized soil (5 kg/ pot).

After that, the inoculum was mixed with

the soil at the rate of 2% (w/w) of soil,

and then irrigated three times a week

before transplanting to ensure even

distribution and growth of each particular

fungus. Other sterilized pots were filled

with sterilized soil and un-inoculated

with the tested fungi were kept as control.

Five transplants were planted in each pot

and a set of three replicates were used for

each particular treatment. The disease

assessment was estimated. Percentages

of infection and diseases severity were

recorded after 120 days from

transplanting date. The arbitrary (0-5)

disease index scale as described by

Bayraktar et al. (2010) was adopted.

Where: 0 = no disease, 1 = 1–20%

diseased, 2 = 21–40% diseased, 3 = 41–

60% diseased, 4 = 61–80% diseased, and

5 = 81–100% diseased. Disease severity

index (DSI) was calculated according to

Ichielevich-Auster et al. (1985) as

follows:

Disease severity index = ∑ (disease severity

scale × numbers of plants at each scale) ⁄

total number of plants.

2.4 Effect of certain biofertilizers on

incidence of root rot disease under

greenhouse conditions

The experiment was carried out in winter

during 2018/2019 and 2019/2020

growing seasons

using five different

types of bio-fertilizers (Table 1) as a

substitution of chemical fertilization

namely; (Cerialien, Biogen, Nitrobein,

Phosphoren and Potassiumag). These

bio-fertilizers were obtained from the

bio-fertilization unit, Agricultural

Research Centre Ministry of Agriculture,

Giza, Egypt as microorganisms in peat

moss carrier substrate. The sterilized pots

which used in this experiment were filled

with the sterilized soil of clay and sand,

(1:1 v/v).

Table 1: Bio-fertilizers used to control root rot diseases of onion.

Bio-fertilizers

Bio- components

Cerialien

Azosperillium sp

Biogen

Azotopacter sp. + Azosperillium sp

Nitrobein

Azotopacter sp. + Azosperillium sp

Phosphoren

As phosphorus solubilizing bacteria at (10

8

cfu/g) Bacillus megaterium var. phosphaticum

Potassiumag

Bacillus verculanes, Bacillus megaterium var. phosphaticum

18

The inoculum of

Fusarium oxysporum

f.sp.

cepae

,

Pyrenochaeta terrestris

and

Sclerotium cepivorum

were added at the

rate of 2% of soil weight and mixed with

the upper surface of the soil, irrigated and

left 7 days for fungal growth. Then the

fresh preparation of each biofertilizer was

mixed separately in the soil at the rates of

(1, 2 and 3 g/kg soil). The transplants of

onion were prepared and immediately

planted in the infested or non-infested

soil with the tested fungi (5

transplants/pot). Three pots were used for

each treatment. Disease incidence was

assessed as the percentages of diseased

transplants as mentioned before.

2.5 Effect of commercial biofungicides

on controlling root rot disease of onion

under greenhouse conditions

Five commercial bio-products,

i.e.

(Bio-

Zeid, Plant guard, Bio-Arc, Rhizo-N and

T34 Biocontrol). Table (2) which

obtained from Organic and Bio-

technology (Sadat city, Menofeia, Egypt)

under Supervision of Biological Control

and Microorganisms Unit, Plant

Pathology Research Institute, Agriculture

Research Center, Giza, Egypt. While,

T34 biocontrol provided by Shoura

Agrochemical Company .These

biofungicides were used to evaluate their

efficiency on controlling root rots of

onion. Three concentrations of each

tested commercial biofungicide

i.e.

1, 2

and 3 g/kg soil were used. Plastic pots

(30 cm in diameter) were filled with

sterilized soil and mixed with the

inoculum of the pathogenic fungi

F.

oxysporum

f.sp .

cepae

, P. terrestris

and

S. cepivorum

at the rate 2% (w/w) of clay

soil, one week before adding

biofungicides and planting, while the

pots were filled with sterilized soil and

infested with the pathogenic fungi only

served as a control. The commercial

biofungicides were added and distributed

in the infested soil at the time of

transplanting. The transplants of onion

were prepared and immediately covered

in the infested or non-infested soil with

the tested fungi (5 transplants /pot).

Three pots were used for each treatment.

Disease incidence was assessed as the

percentages of diseased transplants as

mentioned before. Also, the growth

parameters were recorded.

Table 2: Biofungicides used as bio-agents against the causal pathogens.

Biofungicides

Bio-agents (density/ml)

Bio-Zied

(Trichoderma album),10x10

6

spores/g

Bio-Arc

(Bacillus megaterium) 25x10

6

cell/g

Plant guard

(T. harzianum) 30x10

6

spores/g

Rhizo-N

(B. subtilis) 30x10

6

cell/g

T34 biocontrol

(T. asperellum) 12x10

6

spores/g

2.6 Statistical analysis

Data collected were subjected to the

statistical analysis according to the

standard methods recommended by

Gomez and Gomez (1984) using the

computer program (Costat). The

differences between the mean values of

various treatments were compared by

Fisher’s LSD.

19

3. Results

and Discussion

3.1 Isolation and identification of the

associated fungi with onion root rot

disease

Different fungal isolates

i.e

.

S.

cepivorum

,

F.

oxysporum f.sp.

cepae

,

P.terrestris

,

Botrytis allii

,

Aspergillus

niger

,

F. moniliforme, F.semitectium, F.

proliferatum

,

F. verticillioides

,

F.

anthophilium

and

Penicillium

sp. were

isolated from onion root rots collected

from different Localities in Assiut,

Sohag, Menofeia and Luxor

governorates, Egypt during 2016 and

2017 growing seasons. The fungal

isolates were identified by using the

morphological features of mycelium and

spores as described by Barnet and Hunter

(1986) and Booth (1977)

and confirmed

by Agricultural Botany Department,

Faculty of Agriculture, Al-Azhar

University (Assiut branch), Egypt. Data

in Table (3) indicated that the highest

percentage of occurrence was recorded

by

S. cepivorum

,

F. oxysporum

f. sp.

cepae,

Botrytis allii

and

P. terrestris.

These fungi were the most dominant

fungi in all locations as their frequencies

were 35, 26.3, 10 and 6.6 % respectively.

Also,

Aspergillus niger

,

F. proliferatum

and

F. semitectium

reached it 4 , 3.4 and

3.4 % respectively, followed by

F.

anthophilium,

F. verticillioides

and

F.

moniliforme

3.2 , 3 and 2.9%

respectively, while

Penicillium

sp

exhibited the lowest ones, 2.2 % of the

total count of fungi. These results are in

agreement with those obtained by Ali

(2015) who mentioned that isolated fungi

were, identified as

S. cepivorum

,

A.

niger

,

F. oxysporum

,

P. chrysogenum

,

B.

allii

, and

Py. terrestris

from onion plants

and rhizosphere soil of onion plants from

different locations of Assiut governorate.

Table 3: Occurrence and frequency of root-rot fungi isolated from diseased onion plants

collected from different governorates, Egypt.

Governorate

Location

Isolated fungi

Total

F1

F2

F3

F4

F5

F6

SCL

Py

AS

B.a

Pn

Menofeia

Ashmoun 1

20

5

-

10

5

-

45

Ashmoun 2

5

-

5

15

10

-

5

-

40

Total

25

5

-

5

25

20

5

-

90

Frequency %

29.4

5.8

-

5.8

29.4

18

5.8

-

100

Assiut

AL-Azhar Univ.

10

3

-

10

-

6

5

40

Dronka

40

-

5

-

3

2

50

El-Fath

5

-

38

-

7

-

50

Vegetable markets

3

2

3

2

5

-

20

15

10

5

80

Total

58

5

6

5

-

73

15

26

12

220

Frequency %

26.3

2.2

2.7

2.2

-

33.1

7

11.9

5.4

100

Sohag

Tahta

5

-

5

15

-

5

-

45

Shandaweel

15

-

3

2

20

-

5

-

45

El Munsha

15

3

-

20

-

5

-

43

Total

35

3

5

8

7

55

-

5

10

-

133

Frequency %

26.3

2.2

3.8

6

5.3

41.3

-

3.8

7.5

-

100

Luxor

Armant

10

-

5

10

-

5

-

45

Isna

5

-

20

-

6

-

36

Total

15

5

30

-

11

-

81

Frequency %

18.5

6.2

37

-

13.5

-

100

Total

138

18

16

15

18

17

183

35

20

52

12

524

Frequency %

26.3

3.4

3

2.9

3.4

3.2

35

6.6

4

10

2.2

100

F1: F. oxysporum f.sp. cepae, F2: F. proliferatum, F3: F. verticillioides, F4: F. moniliforme, F5: F. semitectium, F6: F.

anthophilium, SCL: S. cepivorum, Py: Py. Terrestris, AS: Aspergillus niger, Pn: Penicillium sp., B.a: Botrytis allii.

20

The fungus

F. oxysporum

showed the

highest frequency, mean frequency rate,

was 31% followed by

S. cepivorum

(21%) and

A. niger

(20%). While,

P.

terrestris

had the lowest frequency, mean

frequency rate was 5%, followed by

P.

chrysogenum,

and

B. allii

. The

occurrence and frequency of the isolated

fungi were differed from one location to

another. These differences are probably

due to the environmental conditions such

as moisture, temperature and soil type,

dissemination factors of fungi in different

locations and agricultural practices. The

isolated fungi were purified, identified

and the most frequently isolated fungi

i.e.

S. cepivorum

,

F. oxysporum

f.sp

. cepae,

P.terrestris

,

Botrytis allii

and

Aspergillus

niger

were used for further studies.

3.2 Pathogenicity tests

Forty fungal isolates were tested to study

their pathogenic capabilities on onion

plants (Giza 6 Mohassan cv.) under

greenhouse conditions during 2016/2017

growing season. Data presented in Table

(4) illustrated that all tested fungal

isolates were able to infect onion plants

caused root rot diseases with different

degrees of disease severity. Data showed

that

F. oxysporum

f.sp.

cepae

(No. 4)

gave the highest percentage of disease

severity, followed by

P. terrestris

(No.

24) and

S. cepivorum

(No. 15) These

isolates were the most virulent among

all the tested isolates. In which, they

recorded (95.8, 95.7 and 93.4%) disease

severity, respectively. According to

obtained data,

F. oxysporum

f. sp.

cepae

(No. 4),

P. terrestris

(No. 24) and

S.

cepivorum

(No. 15) were selected for

further studies under lab and greenhouse

conditions. These results are in harmony

with those reported by Mahdy et al.

(2018) who found that inoculating onion

bulbs (Giza 20 cv.) with 14 isolates of

Fusarium

indicated that the fourteen

tested isolates were pathogenic to onion

plants.

Table 4: Pathogenicity tests of 40 fungal isolates on onion plants (Giza 6 Mohassan cv.) under greenhouse

conditions during 2016 growing season.

Isolate number

Fungal isolate

Disease Severity (%)

Isolate number

Fungal isolate

Disease Severity (%)

1

F. oxysporum

30

21

Py.terrestris

35.1

2

F. oxysporum

43.3

22

Py.terrestris

86.9

3

F. oxysporum

41.6

23

Py.terrestris

88.2

4

F. oxysporum

95.8

24

Py.terrestris

95.7

5

F. oxysporum

20.2

25

Py.terrestris

19.6

6

F. oxysporum

24.8

26

Py.terrestris

37.1

7

F. oxysporum

53.5

27

Py.terrestris

37.1

8

F. oxysporum

86.2

28

Py.terrestris

56.1

9

F. oxysporum

31.6

39

Py.terrestris

75.1

10

F. oxysporum

24.1

30

Py.terrestris

12.1

11

S. cepivorum

48.5

31

B. allii

10.8

12

S. cepivorum

84.5

32

B. allii

64.5

13

S. cepivorum

89.8

33

B. allii

46.5

14

S. cepivorum

38.8

34

B. allii

32.4

15

S. cepivorum

93.4

35

B. allii

54.4

16

S. cepivorum

23.6

36

A. niger

12.8

17

S. cepivorum

19.7

37

A. niger

12.8

18

S. cepivorum

90.4

38

A. niger

33.4

19

S. cepivorum

32.2

39

A. niger

45

20

S. cepivorum

74.5

40

A. niger

18.9

Un-infested soil

0

LSD at 0.05

3.86

21

As for virulence of each one of the

isolates on bulbs and seedlings of onion,

F. oxysporum

caused severe basal rot and

damping-off as a highly virulent species

were also confirmed by Shalaby et al.

(2013)

who mentioned that screening

trials of the fields infested with onion

white rot disease resulted in eight isolates

of

S. cepivorum

and pathogenicity of

these isolates showed varied degrees

against onion transplants Giza 20 cv.,

ranging from the most aggressive (100%)

in case of isolate Sc2 to the lowest degree

(75%) for isolate Sc1. Strong pathogenic

variations between

S. cepivorum

isolates.

Kafi (2009) who showed that evaluate the

influence of the inoculum sources with

Pyrenochaeta terrestris

inoculum

(transplants and soil) resulted in 100%

infection and significantly reduced the

root development, leaf growth and bulb

weight of cv. Kamlin yellow when

assessed two months after transplanting

and at maturity

.

3.3 Effect of different biofertilizers on

incidence of root rot of onion plants

under greenhouse conditions

Different biofertilizers

i.e.

(Cerialien,

Biogen, Nitrobein, Phosphoren and

Potassiumag) with different

concentrations (1, 2 and 3 g/kg soil) were

evaluated to study theis effectivenes on

incidence of onion root rot diseases

caused by

F. oxysporum

f.sp.

cepae

,

P.

terrestris

and

S. cepivorum

were tested

under greenhouse conditions during

2018/2019 and 2019/2020 growing

seasons. It was clear from data in Table

(5) that all the tested biofertilizers were

effective in reducing the disease severity

of onion root rot disease. The disease

severity decreased as biofertilizers

concentration increased. The treated soil

with different biofertilizers significantly

decreased the disease severity of onion

root rot diseases compared with the

control. Nitrobein, followed by

Phosphoren with all tested concentrations

(1, 2 and 3 g/kg soil) gave the highest

effect in minimizing the disease severity

caused by

F. oxysporum

f.sp.

cepae.

Also, Biogen at 2 and 3 g/Kg soil

revealed higher effects in decreasing

disease severity caused with the same

fungus. Meanwhile, Nitrobein at 3 g/kg

soil was the most effective biofertilizer in

minimizing disease severity, followed by

Phosphoren with the same concentration

during 2019/2020 growing season. On

the other hand, Nitrobein, followed by

Phosphoren each at 2 and 3 g/kg soil

were the best biofertilizers, which

reduced disease severity caused with

S.

cepivorum

. Also, Biogen and Cerialien at

3 g/kg soil revealed the higher effect in

reducing disease severity, while

Potassiumag was less effective in

controlling root rot diseases caused by

S.

cepivorum.

It was shown from the same

Table that Nitrobein, Biogen and

Phosphoren with all tested concentrations

were the most effective biofertilizers in

reducing disease severity caused by

P.

Terrestris

, when applied as soil

treatment. Also, Potassiumag 3 g/kg soil

revealed the highe effect in reducing

disease severity, while Cerialien came in

the last with the same concentrations.

Also, data revealed that the bio-fertilizers

reduced disease incidence caused by the

tested fungi. Under greenhouse

conditions, Nitrobein, Phosphoren and

Biogen was more effective on the tested

fungi. In this respect, Hassouna et al.

(1998), Bhardwaj et al. (2014) and Dhir

(2017) stated that,

Azotobacter

22

brasilensis

and

A. chrococcum

were very

effective against the infection with

R.solani

and

F. oxysporum.

Phosphoren

was effective than Microbin in reducing

pod of peanut. This effect was attributed

to the decrease of population density in

the rhizosphere (Zeidan, 2000). The same

trend was recorded by Emara (2005)

using Rhizobacterin and Phosphoren.

Also, Brown (2012) and Zaghloul et al.

(2007)

observed that

Azotobacter

besides

the N-fixation was able to produce

growth substances and fungal antibiotics,

the response of the crops to the

inoculation could be attributed to the

substances produced by the organisms.

Also, Chung and Wu (2000) recorded the

efficiency of

Bacillus megaterium

var.

phosphaaticum

to control root-rot caused

by

R.solani

and the mycelia growth was

generally reduced, where some isolates

were able to cause a significant reduction

in the damping-off of the plants. Also,

Potassiumag containing

Bacillus

verculanes

was suppressive compared

with the control.

Table 5: Effect of using different biofertilizers on controlling bulb rot of onion diseases

under greenhouse conditions during 2019/2020 growing seasons.

Biofertilizer (A)

Rate of

application

(g/kg soil) (B)

Disease severity (%)

F. oxysporum

Py. Terrestris

S. cepivorum

2019

2020

2019

2020

2019

2020

Cerialien

1

38.5

36.2

34.8

32.4

65.5

62.4

2

30.6

31.1

29.9

30.4

53.7

54.8

3

23.3

24

19.6

19.2

27

25.5

Biogen

1

27.4

30.2

15.8

14.2

35.2

33.8

2

22.1

24.3

10.3

9.6

32.2

31.1

3

12.9

11.5

8.1

7.8

22.8

25.4

Nitrobein

1

18.5

16.2

11.8

14.4

26.6

28.4

2

11.4

10.4

12.7

10.7

19.6

19.2

3

4.6

5

3.5

3.1

15.1

14

Phosphoren

1

22.9

23.1

18.1

17.7

36.5

34.5

2

18.5

19.5

14.5

15

24.1

26.1

3

5.2

5.9

8.5

9.2

17

17.8

Potassiumag

1

35.7

37.6

32.8

28.7

72.1

71.1

2

32.2

34.6

24.8

27.6

67.8

64.2

3

19

21.4

10

9.8

59

47.3

Check

99.65

99.99

94.55

92.14

100

L.S.D. at 5%

A

2.39

0.82

2.23

2.88

2.70

2.78

B

0.59

0.98

1.63

2.59

1.12

A×B

1.44

4.88

2.40

3.99

6.35

2. 74

3.4 Effect of biofertilizers on growth

parameters

The effect of soil treatments with

different biofertilizers

i.e.

(Cerialien,

Biogen, Nitrobein, Phosphoren and

Potassiumag) with different

concentrations (1, 2 and 3 g/kg soil) on

the growth parameters in infected soil

during 2019 and 2020 growing seasons

was studied. Data shown in Table (6) that

the pathogens infested potted plantlets

treated with different biofertilizers

showed significant differences in terms

of Fresh bulb weight, dry bulb weight

and bulb diameter than untreated

plantlets. The maximum Fresh bulb

weight, dry bulb weight and bulb

diameter was recorded using Nitrobein,

Phosphoren and Potassiumag at

23

concentration 3 g/kg soil. From data

presented in the same Table, it could be

noticed that these treatments also

promoted the growth parameters of onion

plants in infested soil with

F. oxysporum

f.sp.

cepae

as compared with the control

treatment during 2019 and 2020 growing

seasons. In this respect, Nitrobein

followed by Phosphoren and Potassiumag

proved to be the most effective in

increasing Fresh bulb weight, dry bulb

weight and bulb diameter as compared

with other treatments and control. As for,

Biogen with all concentrations tested

were found to be the most effective in

increasing the bulb diameter more than

other biofertilizers in infested soil with

F.

oxysporum

f.sp.

cepae

during 2019 and

2020 growing seasons. The same data

Table (6) showed that the highest dosage

of all biofertilizers exhibited higher

increasing of all growth parameters in

infested soil with

P. terrestris

as

compared to their lower dosages and the

check treatment. Nitrobein, Biogen and

Phosphoren recorded higher increasing

of Fresh bulb weight, dry bulb weight

and bulb diameter more than other

biofertilizers in infested soil with

P.

terrestris

. The lowest growth parameters

were observed in infested soil and

untreated with any biofertilizers during

2019 and 2020 growing seasons.

Regarding the effect of biofertilizers on

growth parameters in infested soil with

S.

cepivorum

, highest fresh bulb weight,

dry bulb weight and bulb diameter has

been observed in treated soil with

Phosphoren, Nitrobein and Biogen with

all concentrations tested. However,

Intermediate increases in Fresh bulb

weight, dry bulb weight and bulb

diameter were obtained with Cerialien

and Potassiumag at concentration 3 g/Kg

soil as compared with other biofertilizers

in infested soil with

S. cepivorum

and

untreated ones during 2019 and 2020

growing seasons.

Table 6: Effect of different concentrations of biofertilizers on Fresh bulb weight , dry bulb weight and bulb

diameter of bulb rot of onion Giza 6 Mohassan, cv. under greenhouse conditions during 2018/2019 and 2019/2020

growing seasons.

Biofertilizer

(A)

Rate of

application

(g/kg soil)

(B)

2018/2019

2019/2020

F.

oxysporum

Py.

Terrestris

S.

cepivorum

F.

oxysporum

Py.

Terrestris

S.

cepivorum

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Cerealein

1

22.1

10.3

4.8

31.4

16.2

5.1

37

19.3

5.18

23.3

10.4

4.6

31.1

16.4

5.1

32.3

15.8

5.1

2

31.4

16.2

5.9

47.2

27

5.9

39.7

24.5

6.9

36.8

14.1

6

35.2

25.9

6.2

36.7

23.3

6

3

34.7

16.5

6.3

48.8

37

6.8

49.7

32.3

8.4

35.2

25.9

6.2

50

37

6.6

48.9

33

7.4

Biogeain

1

40.5

24.2

9.1

40.5

23.5

6.6

36.8

23.5

5.1

39.2

23.2

8.8

46.6

28.1

6.7

39.2

24.3

6.2

2

54.8

34.6

9.7

53.8

36.9

6.9

45.1

24.5

6.6

53.3

36.3

10.3

53.3

37.3

7

42.1

28.1

6.6

3

80.1

54.3

11.9

83.9

47.1

9.1

64.2

38.7

9.1

78.7

54

11.6

82.1

46.2

8.8

63

46.2

9.5

Nitrobein

1

53.8

47.2

6.6

50.7

24.8

6.8

40.4

28.5

6.3

50.5

43.8

7.5

48.8

34.7

7.5

39.7

27

6.4

2

83.9

53.2

9.1

54.8

46.9

7

63.1

39.1

8.7

82.3

57.7

7.5

55.2

43.8

7.7

62.5

41.9

8.8

3

102

73.3

10.4

96.1

73.1

10.6

65

44.5

10.3

100

77.1

10.6

98.5

76.1

11.2

65.2

46.2

10.6

Phosphoren

1

36.8

33.5

7.7

38.3

23.2

5.9

54.8

36.9

6.6

48.8

27

6.6

41.7

26.8

6.5

55.3

36.2

6.6

2

65.9

47.1

9.1

51.8

36.2

6.3

83.9

47.1

9.1

64.8

46.2

9.5

48.8

33.8

6.6

82.1

46.2

9.5

3

97.3

71.7

10.3

63.1

39.1

8.7

97.3

71.7

10.7

99.9

73.9

10.4

62.5

41.9

9.5

95.7

76.9

10.9

Potassiumag

1

45

24.8

6.6

36

22.3

5.3

34.6

16.8

4.8

41.6

26.8

6.7

36.9

24.4

5.4

33.1

17.7

4.8

2

63.8

37.3

8.8

39.9

24.8

6.25

38

23

6

60

33

6.6

37

26.8

6.4

38.3

23.3

6.7

3

83.7

57.1

9.1

54.8

38

7.7

47.1

30.2

7.7

82.3

56.2

9.5

53.3

39.3

6.9

37.2

32.5

6.4

Check

16.9

10.3

3.7

10.3

5.2

4

5.9

4.4

2.9

16.9

9.22

4.03

14.1

5.8

3.6

10.1

3.6

2.2

L.S.D. at 5%

A

3.94

2.02

1.46

3.07

2.55

1.45

5.59

3.55

1.81

2.53

2.21

1.46

2.89

1.53

1.51

7.69

3.69

2.16

B

2.59

1.35

0.94

4.09

2.21

1.58

1.54

1.20

1.52

2.11

1.66

1.14

2.63

2.42

1.03

2.99

1.87

1.64

A×B

6.35

3.30

2.31

8.95

6.45

3.49

3.78

2.94

3.73

5.17

4.07

2.79

7.69

7.06

3.02

7.32

4.58

4.02

Such results are in agreement with those

reported by Arfaoui et al

.

(2006), El-

Mohamady and Ahmed (2009) and

Zeidan et al

.

(2012). The enhancement of

24

plant growth parameters and yield

components might be due to ability of

biofertilizers to provide plant by

nutritional requirements and plant growth

regulators and vitamins secured. Isolates

of

Azotobacter

and

Azospirillum

produce

IAA and possessed phosphorus

solublization capability of

R. solani

growth as well as production, ACC

deaminase, siderophore, salicylic acid,

hydrogen cyanide, cellulase, chitinase

and α-1,3-glucanase (Zarrin et al

.,

2009).

3.5 Effect of commercial biofungicides

on control onion root rot diseases

under greenhouse conditions

The efficacy of Rhizo-N, Bio-Arc, Plant-

guard, Biozied and T-34 as biofungicides

on controlling root rot disease of onion

transplants, Giza 6 mohassan cv. was

studied under greenhouse conditions

during 2019 and 2020 growing seasons.

Data presented in Table (7) indicated that

all the tested biofungicides were

effective in reducing the disease severity

of onion root rot diseases. The disease

severity decreased as biofungicidal

concentration increased. The treated soil

with different biofungicides significantly

decreased the disease severity of onion

root rot diseases compared with the

control. T-34 biocontrol at the rate of 2

and 3 g/kg soil was the most effective

biofungicide in minimizing disease

severity caused with

F. oxysporum

f.sp.

cepae

,

P. terrestris

and

S. cepivorum

,

followed by Biozied and Rhizo-N with

the same concentration during 2019 and

2020 growing seasons. These findings

were in agreement with those previously

obtained by Abou-Zied et al

.

(2016) and

El-Naggar et al

.

(2018) reported that

biocides

i.e.

Biozied, BioArc, Plantguard

and Rhizo N were significantly effective

in controlling root rot disease incidence.

Table 7: Effect of different biofungicides on control onion root rot diseases under

greenhouse conditions during 2019/2020 growing seasons.

Biofungicide (A)

Rate of

application

(g/kg soil) (B)

Disease severity %

F. oxysporum

Py. Terrestris

S. cepivorum

2019

2020

2019

2020

2019

2020

Rhizo-N

1

23.8

23.6

17.6

19.5

36.5

36

2

13.8

13.6

9.6

8.9

19.8

19.5

3

5.2

5.9

3.3

4.4

10.7

11.6

Bio-Arc

1

27.4

30.2

15.8

14.5

39.9

38.1

2

16.6

15.2

12.8

11.5

25.7

20.5

3

10.1

10.7

6.5

7.6

13.7

10.7

Plant-guard

1

24.2

24.6

14

15.4

40.7

42.5

2

18.8

19.5

13

13.8

32.2

33.8

3

10.3

11.4

7.4

9.2

15.1

14

Biozied

1

13.8

14.1

9.6

9.2

20.7

21.3

2

9.6

10.4

7.8

8.5

18.8

19.4

3

4.8

5

2.9

3.1

8.1

7

T-34 biocontrol

1

10.7

9.9

4.8

3.3

13.4

12.2

2

5.1

4.3

3.7

3.1

7.8

3

2.2

2.4

1.1

2

6

5.8

Check

99.6

99.9

94.5

92.1

100

L.S.D. at 5%

B

1.95

1.3

2.49

2.41

2.07

2.9

R

0.98

0.81

0.94

1.27

2.03

1.34

B×R

2.41

1.98

2.31

3.13

4.9

3.3

Recently, chemical control is faced with

many difficulties especially what

concerned with their efficacy, selectivity,

toxicity and general impact on the

25

environment (Burrows et al

.,

2007;

Nawar, 2005). As well as the harmful

side effects of the fungicides on human

and environment led to searching new

means or bio-agents with low toxicity

and side effects that can effectively

replace the fungicides in controlling plant

diseases. Therefore, Bio-Arc (Bacillus

megaterium), Bio-Zied (Trichoderma

album) as biocides play a very useful role

as effective and safe means in controlling

root-rots. In this respect, similar results

were obtained by Chavan et al

.

(2004) on

the positive efficacy of treating with bio-

agents, i.e. several strains of bacterial

bio-agents including

Bacillus megaterium

and also by

Trichoderma

spp., to control

damping-off of safflower

.

3.6 Effect of biofungicides on growth

parameters

The effect of soil treatments with

different biofungicides

i.e.

Rhizo-N, Bio-

Arc, Plant guard, Biozied and T34

biocontrol with 3 rates on the growth

parameters in infected soil during 2019

and 2020 growing seasons was studied.

Data in Table (8) illustrated the effect of

different treatments on the Fresh bulb

weight, dry bulb weight and bulb

diameter of Giza 6 mohassan cv. as

compared with the control treatment. All

the biofungicides at three different

concentrations showed the highest

increase of all growth parameters as

compared with the control treatment

during 2019/2020 growing seasons. The

obtained results proved that all

biofungicides with all concentrations

tested significantly increased the growth

parameters as compared to control. In

this concern, T34 biocontrol followed by

Biozied, Rhizo-N, Bio-Arc and Plant-

guard each at 3 g/ kg soil were found to

be the most effective in increase of all

growth parameters of onion transplants

in infested soil with

F.oxysporum

f.sp.

cepae

,

P. terrestris

and

S. cepivorum

as

compared with the control treatment.

Table 8: Effect of biofungicides on Fresh bulb weight, dry bulb weight and bulb diameter of onion plants Giza 6

Mohassan cv. under greenhouse conditions during 2018/2019 and 2019/2020 growing seasons.

Biofungicide

(A)

Rate of

application

(g/kg soil)

(B)

2018/2019

2019/2020

F.

oxysporum

Py.

Terrestris

S.

cepivorum

F.

oxysporum

Py.

Terrestris

S.

cepivorum

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Fresh

Bulb

weight

Dry

Bulb

weight

Bulb

diameter

Rhizo-N

1

53.8

47.2

6.7

38

23.5

5.9

40.4

28.5

5.2

48.8

43.8

6.7

41.8

24.4

5.4

42.1

28.1

6.2

2

67

49.2

9.1

59.3

36.9

8.2

45.1

34.5

6.9

62.3

46.2

9.5

67

36.8

8.9

49.7

35.3

6.9

3

83.9

74.1

10.3

74.8

39.1

8.9

64.2

38.7

9.1

82.3

48.6

10.6

73.3

39.3

9.4

63

44.2

9.5

Bio-Arc

1

43.8

27.7

6.6

36.8

23.3

5.3

37

19.3

5.18

39.2

27.8

6.6

41.6

28.1

5.1

32.2

17.7

5.1

2

57.2

36.9

7.7

54.8

34.8

7.3

39.7

30

6.6

42.2

37.6

6.6

55.3

36.2

6.7

39.3

34.3

6.6

3

74.4

42.9

9.1

63.1

38

8.6

49.7

32.3

8.4

73.5

46.2

9.5

52.5

31.9

8.8

49.2

37.5

8.4

Plant-guard

1

43.6

23.2

4.9

34.6

16.8

4.81

34.6

16.8

4.8

53.8

27

4.8

33.1

17.7

4.8

33.1

15.8

4.8

2

52.1

36.2

6

38

24.5

6.9

38

27.5

6

24

37

6.05

38.3

23.2

6.6

36.7

33.3

6.7

3

58.1

40.4

6.9

47.1

30.2

8.4

47.1

30.2

7.7

61.4

38.2

7.7

47.2

33

7.4

48.9

33

7.4

Biozied

1

66.8

53.5

8.7

50.3

40.2

6.6

54.8

36.9

6.3

60.5

53.2

8.8

45.9

36.8

6.7

55.3

36.2

6.6

2

83.9

57.1

11

63.8

47.2

8.7

63.1

39.1

8.7

80.8

56.3

10.3

69.2

37

9.6

62.5

41.9

8.8

3

87.3

81.7

11.2

83.7

47.1

9.1

64

44.5

10.1

82.4

76.9

11.4

82.3

46.2

9.5

64.2

46.2

9.6

T-34

1

70.5

54.8

9.4

41.8

32.3

7.4

66.8

23.5

6.6

68.6

56.8

9.5

48.8

43.8

9.7

69.2

37

7.6

2

84.8

67.3

10.7

84.8

57.3

9.7

73.9

47.1

9.1

83.3

67.7

10.5

83.3

57.7

9.9

82.3

46.2

9.5

3

95.5

86.3

11.4

102

77.6

10.9

90.3

71.7

10.3

93.3

80.1

11.5

99.8

76.1

11.2

88.8

76.9

10.6

Check

16.9

10.3

3.7

10.3

5.2

4

5.9

4.4

2.9

16.9

9.2

4

14.1

5.8

3.6

10.1

4.3

L.S.D. at 5%

A

3.66

2.94

1.49

2.52

3.19

1.1

5.59

3.55

1.86

2.94

2

1.52

3.22

1.83

1.31

7.69

3.69

2.16

B

1.45

1.43

1.05

1.66

1.84

0.91

2

1.8

0.98

1.98

1.53

0.89

2.13

1.8

0.83

2.15

1.78

0.85

A×B

4.52

3.52

2.58

4.06

4.5

2.24

4.9

4.41

2.41

4.85

3.75

2.19

5.22

4.41

2.04

5.28

4.36

2.1

These findings were in agreement with

those previously obtained by Abou-Zied

et al

.

(2016) and El-Naggar et al

.

(2018).

reported that biocides

i.e.

Biozied,

26

BioArc, Plantguard, Rhizo N and clean

root that reflect on the increase in plant

height, bulb diameter, bulb weight and

total bulb yield compared with untreated

after 90 days from sowing. Plant growth

promoting Rhizobacteria can promote

plant growth and development either

directly and indirectly. Direct stimulation

includes biological nitrogen fixation,

producing phytohormones like auxins,

cytokinines and gibberellins, solubilizing

minerals like phosphorus and iron,

production of enzyme and induction of

systemic resistance. While, indirect

stimulation is basically related to

biocontrol, including antibiotic

production chelation of available Fe in

the rhizosphere, synthesis of extracellular

enzymes to hydrolyze the fungal cell wall

and competition for niches within

rhizosphere (Castro et al., 2009; Vanloon,

2007; Zahir et al., 2004). Generally,

growth promotion resulted by the

biocontrol agents which may be due to

antagonistic fungi and bacteria in plant

root zone.Root zone antagonistic fungi

and bacteria are able to generate a wide

array of secondary metabolites which can

have a positive influence on plant

growth, enhancing the availability of

minerals nutrients, improving nitrogen

fixation, decreasing susceptibility to frost

damage, improving plant health through

the biocontrol of phytopathogens,

inducing systemic plant resistance and

facilitating plant establishment growth

and development.

References

Abou-Zied N, Mahmoud N, Saleh R, 2016.

Effect of some biotic and abiotic

applications on control of fusarium wilt

of pepper plants. Egyptian Journal of

Phytopathology 44(2): 103–118.

Ali TIH, 2015. Microbiological control of

certain plant diseases.Ph.D.Thesis,

Microbiology Department, Faculty of

Agriculture, Mini University, Egypt, 10

pp.

Arfaoui A, Sifi B, Boudabous A, El

Hadrami I, Chérif M, 2006.

Identification of Rhizobium isolates

possessing antagonistic activity against

Fusarium oxysporum f.sp. ciceris, the

causal agent of Fusarium wilt of

chickpea. Journal of Plant Pathology

88(1): 67–7.

Barnett HL, Hunter BB, 1986.Illustrated

genera of imperfect fungi, 4

th

ed.

Macmillan Publishing Co, New York,

USA, 218 pp.

Bayraktar H, Türkkan M, Dolar FS, 2010.

Characterization of Fusarium

oxysporum f.sp. cepae from onion in

Turkey based on vegetative

compatibility and rDNA RFLP analysis.

Journal of Phytopathology 158(10):

691–697.

Bhardwaj D, Ansari MW, Sahoo RK, Tuteja

N, 2014. Biofertilizers function as key

player in sustainable agriculture by

improving soil fertility. plant tolerance

and crop productivity. Microbial cell

factories 13(1): 1–10.

Booth C, 1977. Fusarium laboratory guide

to the identification of the major

species”, Commonwealth Mycological

Institute, Kew, Surrey, England, 58 pp.

Browen M, 2012. Population of Azotobater

in rhizosphere and effect of artificial

Inoculation. Plant and Soil 17(3): 15.

Burrows F, Louime C, Abazinge M,

Onokpise O, 2007. Extraction and

27

evaluation of chitosan from crab

exoskeleton as a seed fungicide and plant

growth enhancer. merican-Eurasian

Journal of Agricultural & Environmental

Sciences 2(2): 103–111.

Castro RO, Cornejo HAC, Rodriguez LM,

Bucio JL, 2009. The role of microbial

signals in plant growth and development.

Plant Signal Behav 4(8):701–712.

Chavan RA, Bharose AA, Pawar RB, Patil

VD, 2004. Screening of safflower

germplasm lines and penetration

mechanism of Trichoderma virdie

against Fusarium wilt. Journal of Soils

and Crops 14(1): 79–82.

Chung IY, Wu WS, 2000. Effect of Bacillus

megaterium. Plant Pathology Bulletin

9(2): 59–68.

Conn K, Lutton J, Rosenberger S, 2012.

Seminis Vegetable Seeds. Inc. Plant

Health.

Dhir B, 2017. Bio-fertilizers and bio-

pesticides: Eco-friendly Biological

Agents. In Advances in Environmental

Biotechnology, Springer, Singapore,

167–188 pp.

El-Mohamedy RSR, Ahmed MA, 2009.

Effect of biofertilizers and humic acid on

control of dry root rot disease and

improvement yield quality of mandarin

(Citrus reticulate Blanco). Research

Journal of Agricultural and Biological

Science 5(2): 127–137.

El-Naggar MAA, Zaki MF, El-Shawadfy

MA, 2018. Management of onion root-

rot diseases caused by soil born fungi

under Middle Sinai conditions. Middle

East Journal of Applied Sciences 8(1):

91–99.

Emara DA, 2005. Integrated management of

some root rot diseases of Pelargonium

graveolens L. Ph.D. Thesis, Faculty of

Agriculture, Cairo University, Egypt.

FAO, 2017. Onion (dried) production in

2017: crops world regions production

quantity from pick lists. Food and

Agriculture Organization, Statistics

Division (FAOSTAT 2018),

https://www.fao.org/ faostat/en/#data/QC,

Accessed 18 July 2019.

FAO, 2018. Food and Agriculture

Organization of the United Nations.

Rome, Italy, 57 pp.

Gomez KA, Gomez AA, 1984. Statistical

procedures for agricultural research.

Second edition, John Wiley and Sons,

Inc., New York, USA, 680 pp.

Hassouna MG, El-Saedy MAM, Saleh HM,

1998. Bio-control of soil-borne plant

pathogens attacking cucumber (Cucumis

sativus) by rhizobacteria in a semiarid

environment. Arid Land Research and

Management 12(4): 345–357.

Hossain M, Ahmed M, Ehsanul Haq M, Al

Maruf M, Nabil MN, 2017. Quality

seed of onion: effect of micro and

macronutrients. Annual Research &

Review in Biology 20: 1–11.

Ichielevich-Auster M, Sneh B, Koltin Y,

Barash I,1985. Suppression of damping-

off caused by Rhizoctonia species by a

nonpathogenic isolate of R. solani.

Journal of Phytopathology 75: 1080–

1084.

Kafi AMD, 2009. Influence of inoculum

sources of Pyrenochaeta terrestris on

pink root disease development, growth

and bulb yield of onion. Ph.D. Thesis,

Crop Protection Department, Faculty of

Agriculture, University of Khartoum,

Sudan.

28

Khan SA, Jameel M, Kanwal S, Shahid S,

2017. Medicinal importance of Allium

species: a current review. International

Journal of Pharmaceutical Sciences and

Research 2 :29–39.

Mahdy HA, Eisa NA, Khaled EE, Khalifa

MMA, Gamal AA, 2018. Identification

of Fusarium species causing onion basal

rot in Egypt and their virulence on seeds,

seedlings and onion bulbs. Annals of

Agricultural Science, Moshtohor 56(1):

79–88.

Marrelli M, Amodeo V, Statti G, Conforti F,

2019. Biological properties and

bioactive components of Allium cepa L.:

focus on potential benefts in the

treatment of obesity and related

comorbidities. Molecules 24: 1–18

Nawar LS, 2005. Chitosan and three

Trichoderma Spp. To control Fusarium

crown and root rot of tomato in Jeddah,

Kingdom of Saudi Arabia. Egyptian

Journal of Phytopathology 33 (1): 45–58.

Shalaby ME, Ghoniem KE, El-Diehi MA,

2013. Biological and fungicidal

antagonism of Sclerotium cepivorum for

controlling onion white rot disease.

Annals of Microbiology 63(4): 1579–

1589.

Tjamos EC, Tjamos SE, Antoniou PP,

2010. Biological management of plant

diseases: highlights on research and

application. Journal of Plant Pathology

92: 17–21.

Vanloon LC, 2007. Plant responses to plant

growth-promoting rhizobacteria.

European Journal of Plant Pathology

119: 243–254.

Yang W, Kim J, Lee JY, Kim C, Hwang C,

2019. Antihyperlipidemic and

antioxidative potentials of onion (Allium

cepa L.) extract fermented with a novel

Lactobacillus casei HD-010. Evidence-

Based Complementary and Alternative

Medicine 2019: 1–10.

Zaghloul RA, Hanafy EA, Neweigy NA,

Khalifa NA, 2007. Application of

biofertilization and biological control for

tomato production. In 12

th

Conference

of Microbiology, 18–22 pp.

Zahir ZA, Muhammad A, Frankenberger

WT, 2004. Plant growth promoting

rhizobacteria: applications and

perspectives in agriculture. Advances in

Agronomy 81: 97–168.

Zarrin F, Saleemi M, Muhammad Z, Sultan

T, Aslam M, Chaudhary MF, 2009.

Antifungal activity of plant growth-

promoting rhizobacteria isolates against

Rhizoctonia solani in wheat. African

Journal of Biotechnology 8(2): 219–225.

Zeidan EH, 2000. Soil treatment with bio-

fertilizers for controlling peanut and pod

rot diseases in Nubaria Province.

Egyptian Journal of Phytopathology 28:

17–26.

Ziedan EH, Mostafa HM, Elewa IS, 2012.

Effect of bacterial inocula on Fusarium

oxysporum f.sp. sesame and their

pathological potential on sesame.

Journal of Agricultural Technology 8(2):

699–709.