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ISSN 2278-2540 | DOI: 10.51583/IJLTEMAS | Volume XV, Issue VI, June 2026
Biological Control of Meloidogyne incognita using Mycorrhizal
Fungi in Central Sudan
Elnour, A. E.
1
, O.O. Elbashir
2
and G. A .A. Elbadri
3
1
Gezira Research Station, Agricultural Research Corporation, Pant Pathology section, Wad Medani
Sudan.
2
Geziera University, Plant Pathology Center, Wad Medani, Sudan.
3
Agricultural Research Corporation, Crop Protection Research Center. Plant Pathology section, Wad
Medani Sudan. Current address Agriculture house Company, Riyadh, KSA.
DOI:
https://doi.org/10.51583/IJLTEMAS.2026.150600074
Received: 14 June 2026; Accepted: 19 June 2026; Published: 06 July 2026
ABSTRACT
There are many pests and diseases damaging both the quality and quantity of tomato production. Plant
parasitic nematodes are one of them. They represent an important constraint on the delivery of global food
security. Damage caused by plant-parasitic nematodes has been estimated at US$ 80 billion per year. Effects
of three concentrations of mycorrhizal fungi were studied for control of Root Knot Nematode. Different
tomato accessions were screened for Root-Knot Nematode susceptibility. All the tested concentrations of
mycorrhizal fungi on nematode infected plants were produce positive effects i.e. nematode numbers were
highly decreased compare to control which also appear as improvement of the growth of infected plants at
various degrees. Among three tested concentrations, generally, middle concentration 5 ml of mycorrhiza is
most effective followed by high one 6 ml of mycorrhizea and the lowest effect produced by lower
concentration 4 ml of mycorrhizea. Mycorrhizea fungi appear as most effective control measures of root-knot
nematode in tomato.
INTRODUCTION
Plant parasitic nematodes cause loss of 10-% of the whole loss in the crop causing billions of dollars of crop
losses across the globe annually. (Cuvaca et al., 2026, Rady et al., 2021 ). Even this however, is an under
estimate as nematode damage can be easily overlooked. Root knot nematode is sedentary endoparasites that
induce the formation giant cells in the roots, from which nematode feed to complete its life cycle (Baldacci-
Cresp et al., 2015, Jaouannet and Rosso 2013). The availability of water and nutrients to the plant decreases
while the giant cells are located close to the root systems xylem and phloem. (Siddqui et al., 2014).
Morphological and biochemical alterations induced by nematode parasitism cause abnormal growth of plants,
nutrient deficiency, symptoms, roots with galls, forking and other deformations (Palomares-Rius , 2017,
Moens et al., 2009). It is estimated that total annual crop losses of 14.6% is caused by plant-parasitic nematode
in tropical and sub-tropical climates (Nicol et al., 2011). Annual worldwide agricultural losses caused by
plant-parasitic nematodes are estimated to be $137 billion with $13 billion in the United States (Elling, 2013,
Howland and Quintanilla, 2023). Chemical control remains the predominant strategy among available
approaches for nematode management. In recent years, a new generation of synthetic nematicides with distinct
biochemical targets and improved selectivity has emerged (Yan et al. 2026 ). Health and hazards concerns
which have elicited close security by regulatory agencies resulted in increasing restriction or prohibition of
use nematicides (Kumar and Donthi, 2024, Osman and Viglierchio, 1998). Natural products seem to avoid
environmental problems caused by synthetic pesticides and many researchers are trying to identify effective
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natural products to replace synthetic pesticides (Adam et al. 2024, Mfarrej and Rara, 2019, Kim et al., 2005).
The high cost and inconsistent effect of synthetic nematicide, together with their scarcity and ill- effect on an
environment and public health, have increased augmented interest on alternative methods of the management
for plant parasitic nematodes (Abd-Elgawad 2021, Viaene and Abawai, 1998).
There are many pests and
diseases damaging both the quality and quantity of tomato production. Plant parasitic nematodes are one of
them. They represent an important constraint on the delivery of global food security. Damage caused by plant-
parasitic nematodes has been estimated at US$ 80 billion per year (Nicol et al.,2011). Numerous studies have
been conducted to determine the damage potential of Meloidogyne species on several vegetable crops
including tomato, and different management strategies have been proposed. With the phase-out of methyl
bromide, in particular, the problem of Meloidogyne spp. on tomato gained new inters (Seid et al., 2015).
However, these were not compiled and presented in a way to help different stakeholders. Thus, the objective
of this study was controlling root-knot nematodes by using Mycorrhizea fungi.
MATERIAL AND METHODS
Nematode sample collection
Root samples of tomato, eggplant and okra showing typical symptoms of root knot nematode were collected
from 3 states in Central Sudan Geziera (Bida, Fadasi, and Dinagela), and Sinar (Grisli, Aradiba and Kageak),
Kharoum (Shambat, Elgili, Wad Ramli and Elbambonab), (Table1). Collect samples were brought to the
laboratory to extracts the causal nematode.
A total of 3-5 roots from each plant were selected and cutting roots .Then, the infected roots were put into
polythene bags, labeled and brought to the laboratory of plant pathology in Agricultural Research Corporation.
All the samples after investigation were found to be Meloidogyne incognita since this species was found the
most dominant in Sudan(Yassin, 1986).
Table (1). Roots samples collected from different location for females’ extraction:
State
Location
Plant species (local
name)
Scientific name
Gezira
Bida
Egg plant
Solanum melongena
Eldnagila
Okra
Abelmoschus
esculentus
Fadasi
Egg plant
Solanum melongena
Sennar
Grisli
Tomoto
Solanum tubersoum
Aradiba
Tomoto
Solanum lycopersicum
Kageak
Okra
Abelmoschus
esculentus
Khartoum
Shambat
Tomato
Solanum lycopersicum
Elgili
Egg plant
Solanum melongena
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Wad Ramli
Egg plant
Solanum melongena
Elbanbonab
Tomato
Solanum lycopersicum
The effect of Mycorrhizea on the nematode infection
Mycorrhizea treated on healthy tomato plants previously infected with root knot nematode M. incognita
extractions to assess the efficacy of mycorrhizea on reduction of nematode infection. To assess the effect of
mycorrhizea on nematode infection, disease rating index, number of eggs, galls of nematode. Number of
colonies of mycorrhizea. Fresh and dry Weight of shoot and root and length of infect root were recorded.
Nematode extractions
Nematode eggs were extracted from heavily galled roots as described by (Kim et al., 2001). Roots were
washed, cut into 1-2cm segments and maceration in blender for 20 seconds. The macerate was filled into a
flask containing 500ml of 1.25% Sodium hypochlorite (Naocl), solution and shaken for 3 minutes to liberate
eggs from the gelatinous matrix. Eggs were separated from plant debris by passing the egg suspension
successively through sieves of 200μm, 100μm and 25μm mesh size, remove excess chlorine, the eggs from
the 25μm sieve were washed several times with tap water. Eggs from the 25μm sieve were then collected in
tap water. To confirm the pathogenicity of the extracted nematode (egg) inoculation tomato is needed.
Preparation of tested plants
Seeds of tomato variety Beto-86 were maintained in polyethylene bags and stored at 4ºC. They were surface
sterilized using 1.25 % Sodium hypochlorite for 20 minutes and rinsed thoroughly in sterile distilled water
and were grown in trays in commercial peat-based substrate. When plants reached the 4-5 leaf stage they were
transplanted into 2:1 mixture of clay and sand sterilized soil. Plants were watered adequately throughout the
experiment period. Tested plants were inoculated at three weeks old after transplanting.
For inoculation, the eggs were suspended in tap water the total number of eggs per ml were around.
Experiments were conducted in a greenhouse in Geziera Research Station Wad Madani, Sudan. The
treatments were arranged in a complete randomized block design with three replicates. Observations of the
infected plants and rating is done as follows.
Nematode Rating Index
The nematode gall rating index was rated on a 0 – 10 scale with 0 = n galls and 10 = completely galled (
Salazar, and Schroeder, 2018 ). Numbers of galls and eggs masses induced were counted under microscope.
For the counting of eggs roots were immersed in an aqueous solution of phloxin B (0.15g/1) for 15 min to
stain the egg masses. The number of root galls per root system was counted and prevalence determined on the
fooling scale: 0=0.1=1-2.2=3-10.3=11-30, 4=31-100 and 5= greater than 100 galls. The number of egg masses
per root system was counted and prevalence determined on the following scale: 0=0.1=1-2.2=3-10.3=11-30,
4=31-100 and 5= greater than 100 egg masses (Tylor and Sasser, 1978).
At the end of experiments, the plant fresh, dry shoot and root weight, number of galls and eggs per plant and
galling index was scored.
The experiment was conducted two times at different seasons. Experiments were conducted in Random
Complete Block Design (RCBD). Data were then analyzed using Mstat program. Treatment means were
separated using Duncan’s multiple range tests. Statistical differences referred to in the text are significant at
P<0.05.
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Arubecular Mychorrhizal fungi (AMF)
Collection, isolation, and counting of fungi spore
Mycorrhizea spp which used during this investigation was isolated from grasses (Cynodon dactylon) in Gezira
research station, Wad Medani, using sieves with different size from (250 μm, 180 μm, 63μm and 45μm).
Using a fine ruler, stereomicroscope, diameter of the ocular field is 4x magnifications for a comfortable field
of view (diameter 5 mm). The area of the field was 19.6 mm (Radius = 2.5 mm). Using plastic petri dishes
for counting, enough water added to have complete coverage of the base, diameter (85 mm), area of the base
calculated 5672 mm. from this datum, and the area of the ocular field, total number field in the dish is : 5672/
19.6= 289. Extract spores and use sieves to separate out as much root and organic detritus, added the spore
suspension to a petri dish and then randomly rotate the dish to spread out the spores, then place spore
extraction in a test tube, dilute 1:1 with water while overtaxing, remove specific volume (10 ml) transfer to
petri dish, and recount.(Giovanetti et al., 1980).
Inoculum preparation
Inoculation of tomato seedling with mycorrhizal fungi was carried out according to (Prsanthi et al., 2016), in
which 4 ml (1000 spores), 5 ml (1250 spores) and 6 ml (1500 spores) means 250 spores /ml were placed 5 cm
below surface soil after transplanting. Different parameters were asses to explain the effect of mycorrhizea
on nematode infection these parameters include.
Plant root length and Mycorrhizal root lengths
In the experiment, all of the roots of an experimental plant collected and measured and then grid-line intersect
method used to measure mycorrhizal root length, the roots colonized by mycorrhizal fungi (Giovanetti, et al.,
1980). (cover table to percentage colonization when divided by root length), and the total length of roots
produced by the plant. Several small samples are removed from the main root mass after blotting, pooled, and
fresh weight is measured (FW1), the remaining roots of the sample are weighted (FW2), staining root sample
in 0.05% ink , place in paper envelopes and dry for at least 48 hr. at 62C. Measure dry weight of large sample
(DW2) (Giovanetti et al., 1980).
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Spread roots in plastic petri dish in which a grid with 0.5*0.5
A square is affixed to the base. Collecting the following data:
(i)Total number of intersects between lines and roots (R1).
(ii) Total number of intersects where the root is mycorrhizal (R2).
The data above data is used to calculate plant root length and mycorrhizal root length of the total sample based
on the assumption that distribution of mycorrhizae in the small pooled sample reflects the distribution of
mychorrhize throughout the entire root system.
Dry weight of sample stained to measure colonization (DW1)
This calculation is performed setting up a proportion and solving for x (DW1), as follows:
DW2/FW1= x/FW1
(FW2)(x)= (FW1) (DW2)
X or DW1= (FW1) (DW2)/FW2
Mycorrhizal Root Length in Large (Unstained) sample (R3)
This calculation is an estimate on the actual measurement of mycorrhizal root length in the small stained
sample (R2). Another proportion is set up and solved for x (R3). As follows:
R2/DW1=x/DW2
(DW1)(x)= (R2)(DW2)
X or R3= (R2) (DW1)/ DW1
Total Mycorrhizal Root Length = R2+R3
Total plant root length can be estimated by substituting R1for R2 in the same set of calculation. Method,
(Giovanetti et al., 1980)
Stained Roots were spread in a plastic petri dish in which a grid with 1×1 cm squares was affixed to the base,
and the following data was Collected :
(i) total number of intersects between lines (Vertically V and horizontally H ) = roots(R
1
)
(ii) total number of intersects where the root is mycorrhizal (R
2
)
Then R
2
/R
1
= % of AMF colonization in root.
RESULTS
Effect of Mycorrhizae in root-Knot Nematode infection
All the tested concentrations of mycorrhizal fungi (4, 5 and 6 ml) were negatively affected Meloidogyne
incognita culture compared to the control. The reactions against tested nematode varied from one
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concentration to another. Generally, the mycorrhiza inhibited the nematode forming galls and the nematode
number of infected plants. (Table 2).
Table (2). Infection %, shoots and roots fresh and dry weight of nematode infected plants with three
concentrations of mycorrhizea.
Concentration
of
mycorrhizae
Fresh weight of
shoots
Dry weight
of shoots
Fresh weight
of roots
Dry weight of roots
4 ml
19.12a
2.867a
5.922a
1.133b
5 ml
19.59a
2.922a
6.267a
1.278b
6 ml
19.23a
2.911a
5.789a
1.133b
control
16.94b
2.067b
6.611a
2.000a
Cv%
5.96
15.0
14.63
18.39
S.E±
1.243
0.399
0.809
0.065
Values in the same Colum followed by different letters significantly different according to Tukeys multiple
range tests (p< 0.05).
Concentration of
mycorrhizae
Colonization of
mycorrhizae
Nematode
percentage
Mycorrh
izae root
length
Total
plant root
length
Number
of galls/
plant
Number
of eggs/
plant
1 ml
62.91b
34.71b
10.81a
15.51a
620.6 b
290.9 b
2 ml
89.24a
34.82b
10.71a
16.03a
600.9 b
275.9 b
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3 ml
73.42b
36.21b
9.578a
13.859a
590.5 b
280.7 b
control
0.000c
62.48a
0.000b
1.85 b
734.7 a
328.5 a
Cv%
13.03
8.7
31.0
29.5
30.1
25.2
S.E±
4.419
13.36
2.362
3.465
2.892
1.683
Table (3). Number of mycorrhizea colonies, percentage Root-knot nematodes, mycorrhizea root length,
and total length plant roots and number of galls eggs / plant root.
Values in the same Colum followed by different letters significantly different according to Tukeys multiple
range tests (p< 0.05).
All the tested concentrations of mycorrhizal fungi (4, 5 and 6 ml) were negatively affected Meloidogyne spp
compare to the control. The reactions against tested nematode varied from one concentration to another.
Generally, the mycorrhiza inhibited the nematode forming galls and the nematode number of infected plants.
All concentrations of mycorrhizea (4, 5 and 6 ml) used showed significant reduction on the infection by
nematode during 2015- 2016. All parameters measured here were show significant difference compare to
untreated plants. These parameters include rating index, fresh and dry shoot and root and total root length,
colonization of both nematode and maycorizae and numbers of eggs and galls. which recorded increasing in
the control reaching 9.000 compare to 4.000 for treated. Mycorhizae produced increasing the fresh weight of
shoots from 19.23g to 16.94. significant increasing in shoots dry weight 2.86g which was higher than control
plants recorded 2.0g only, fresh and dry root weight of treated plants were decreased from 5.7g to 1.1g while
increased in the control 6.6g – 2g (table 2), mycorrhizea colonization showed significant increasing in treated
plants reached 89%.Compare to zero in control, nematode percentages significantly reductions on mycorrizal
plant reached62%, while control recorded 34%.for treated plants were found between 34 to 43% compared to
the control which 62%, total numbers of plant roots containing mycorrhizea were found10 cm. However, the
treated plants length 16 cm compared to the control plants length 8 cm, number of eggs showed significant
reduction reached 590 egg/plant while increasing in control 734.7, number of galls showed reduction in
forming galls reached 275/plant compare to increasing in control 328.5/plant (table 3).
Table (4). Infection %, Fresh and Dry weight shoots and roots of the infection plants with three
concentration of mycorrhizea Second season.
Concentration of
mycorrhizae
Infection
percentage
Fresh weight of
shoots
Dry weight of
shoots
Fresh weight of
roots
Dry weight of
roots
1ml
3.889a
20.34a
3.067a
6.289b
1.546b
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2ml
4.000b
21.10a
3.400a
6.267 ab
1.493b
3ml
3.778b
20.73a
2.756ab
6.367b
1.667b
control
9.333a
18.00b
2.144b
7.556a
2.411a
Cv%
21.28
6.15
24.99
16.90
26.15
S.E±
1.248
1.518
0.504
1.280
0.217
Values in the same Colum followed by different letters significantly different according to Tukeys multiple
range tests (p< 0.05).
Table (5). Number of mycorrhizea colonies, percentage root-knot nematodes, mycorrhizea root length, total
length of the plant roots and number of galls & eggs / plant root, second season.
Concentration
of
mycorrhizae
Colonization
of
mycorrhizae
Nematode
percentage
Mycorrhiz
ae root
length
Plant root
length
Number of
galls / plant
Number
of
eggs/pla
nt
1ml
69.26c
38.32b
10.63a
15.99a
590.8 b
299.9 b
2ml
90.93b
38.60b
12.87a
19,04a
600.7 b
280.6 b
3ml
75.96b
39.11b
12.44a
17.20a
580.5 b
295.0 b
control
0.000a
63.20a
0.222b
3.00b
650.8 a
390.9 a
Cv%
15.66
8.14
25.98
28.31
29.5
30.5
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S.E±
61.415
13. 362
5.514
13.670
3.465
3.627
Values in the same Colum followed by different letters significantly different according to Tukeys multiple
range tests (p< 0.05).
During this season all treated plants showed significant reduction in gall formation and infection by nematode
ranging from 1- 4 compared to high recorded for control which reached 9, for both fresh and dry weight were
significantly increased compere to control, i.e. fresh weight reach 18 g while treated plants showed 21 g, dry
weight was increased3.4g for the treated plants while decreased in untreated plants weight 1- 2g,fresh and dry
weight of the treated plants roots showed significant reduction in their weight compared to the control roots
weight, colonization of mycorrhizea was showed increasing in treated plants retched 90 compared to the
control zero, nematode percentage was showed significant reduction in treated plants reached 38% compare
to control 63%. The total length was increased in the treated plants (17 -15 cm) compared to the control plants
which was only 3 cm (Table, 2).
DISCUSSION
A number of genera and species of nematodes are highly damaging to a great range of hosts, including foliage
plants, agronomic and vegetable crops, fruit and nut trees, turfgrass, and forest trees (Williams et al., 2017).
Tomato is most important vegetable crop worldwide. Meloidogyne as parasitic nematode among restricting
tomato production factor regarding their damage of the root system inflicting infection and deform the
function of the normal root system to absorb water and nutrients, in addition they help soil borne pathogen to
invade the plant root system. Continued monocropping practiced by commercial farmers in protected
greenhouses (Matlala et al., 2025). provides the ideal environment for crop growth but makes it more
vulnerable to infection by plant-parasitic nematodes. Many control measures were used to minimize the losses
caused by such nematode like e.g. Rotation, Resistant Varieties, Cultural and Management Practices and
nematicdes. Each of these measures shows degree of control with many disadvantages. Using of natural
component as control agent is appear more safety unrespect of their names applied ranging from biological
to natural control. Combination of plant inoculation with a commercial mycorrhizal formulation with half or
full fertilizer application rates was evaluated for the effects on plant growth and yield and mycorrhization
occurrence throughout two consecutive field tomato crops in southern Italy (Candido, et al.,2013). Arbuscular
mycorrhizal fungi (AMF) are obligate root symbionts that can protect their host plant against biotic stress
factors such as plant-parasitic nematode (PPN) infection (Schouteden et al., 2015) . The AMF symbiosis also
leads to an altered root exudation composition and level (Hodge, 2000; Jones et al., 2004), which can in turn
impact the PPN in the rhizosphere in terms of hatching, motility, chemotaxis, and host location (Schouteden
et al., 2015). In our experimental data mycorrchizae reveal very good enhance of treated tomato plans through
both decreasing the rate of infection and improve the plant growth. Inoculation of the plants in the nursery
with AMF allows sufficient time to establish the symbiosis before transplanting and prior to pathogen
exposure same result were founded by (Nerva et al., 2023 Talavera et al. 2001). The seedlings of tomato were
positively response to association with mycorrhizae, showing growth increment, these results reinforced the
previous studies with the same combination of host and AMF Anjos et al., (2005). The decrease in plant shoot
development associated with nematode parasitism usually is related to the interruption of water and nutrients
translocation by the giant cells same as Cofcewicz et al., (2001). The nutritional benefit promoted by
mycorrhizea on tomato plants contributed for increasing the resistance or tolerance to M. javanica m. javanica
same as found by Cofcewicz et al., (2001). The establishment of mycorrhizal prior to nematode contact was
beneficial for tomato plants conferring conditions for improved plant growth in the presence of the pathogen.
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