Biocontrol Science, 2013, Vol. 18, No.

3, 117−127

Review

Integrated Management Strategies for

Tomato Fusarium Wilt
CAROLINE F. AJILOGBA, AND OLUBUKOLA O. BABALOLA＊

Department of Biological Sciences, Faculty of Agriculture, Science and Technology

North-West University, Mafikeng Campus, Private Bag X2046 Mmabatho 2735

Received 20 December, 2012/Accepted 30 January, 2013

Fusarium wilt is caused by the fungal pathogens, Fusarium oxysporum or Fusarium solani.
It is a devastating disease that affects many important food and vegetable crops and a major
source of loss to farmers worldwide. Initial strategies developed to combat this devastating
plant disease include the use of cultural, physical and chemical control. None of these strate-
gies have been able to give the best results of completely ameliorating the situation except for
the cultural method which is mainly preventive. A good knowledge of the nature, behaviour and
environmental conditions of growth of the disease agent is very important to controlling the
disease development in that case. Biological control has been shown to be an environmentally
friendly alternative. It makes use of rhizospheric and endophytic microorganisms that can sur-
vive and compete favourably well with the Fusarium wilt pathogen. They include plant growth-
promoting rhizobacteria（PGPR）such as Bacillus spp. and Pseudomonas spp.. For PGPR to
control or inhibit the growth of the Fusarium wilt pathogen, they make use of mechanisms such
as indole acetic acid production, siderophore production, phosphate solublilization, systemic
resistance induction and antifungal volatile production among others.

Key words：Biological control / Disease development / Pathogen / Plant growth-promoting rhizobacteria.

INTRODUCTION
Fusarium is one of the more troublesome genera of
fungal plant pathogens, causing devastating diseases
like Fusarium wilt and Fusarium root/stem rot in numer-
ous economically important crops. It is one of the most
serious diseases affecting tomato plants throughout the
world, especially in upland areas（Charoenporn et al.,
2010） . Fusarium wilt is caused by Fusarium oxysporum
Schlecht or Fusarium solani. Formae speciales of F.
oxysporum can usually only cause Fusarium vascular
wilt on one plant host species. The predominant hosts
for F. solani are vegetable crops but some strains
may be infectious to man. There are more than 100
Fusarium vascular wilt diseases worldwide（Burgess et
al., 2008）. Apart from causing diseases, they colonize
outer cells of roots as harmless endophytes after the
pathogen has killed the root tissues and others live as
saprophytes in soil（Burgess et al., 2008） . Some strains
of F.oxysporum are not pathogenic and may even
antagonize the growth of pathogenic strains and can be
used as biological control agents（Fravel et al., 2003）.
To provide an environmentally friendly Fusarium disease
control system, the use of antagonistic microorganisms
represents an alternative disease management strategy
（Lugtenberg and Kamilova, 2009）.
This review will summarize the knowledge available
for the use of native Bacillus spp. with biocontrol
potential to combat Fusarium wilt of tomatoes. The
origin, taxonomy and morphology of the Fusarium wilt
pathogen will be introduced together with the pathogen
and the disease. The initial management strategies will
be reviewed and then the use of biological control will
be discussed.

＊
Corresponding author. Tel（work）: +27-183892568, Fax: FUSARIUM WILT AND THE FUNGAL
+27-183892134, E-mail: olubukola.babalola（a）nwu.ac.za PATHOGENS
 118 C. AJILOGBA ET AL.
Origin, taxonomy and morphology of Fusarium
F. solani comprise a complex group of species that
are widely distributed worldwide in soils. It causes
tuber, root, and stem rots of plants. It includes at least
50 subspecies lineages（Schollenberger et al., 2005）.
Taxonomically F. solani belongs to the kingdom Fungi,
phylum Ascomycota, subphylum Pezizomycota, class
Sordariomycetes, subclass Hypocromycetidae, order
Hypocreales, family Nectriaceae and genus Fusarium
（Fry, 2004） .
Distribution and environmental conditions
Weather conditions such as temperature and humid-
ity normally affect the distribution of F. solani（Kosiak et
al., 2004） . Fusarium spp. is found to be widely distrib-
uted in nature in various environments. Toxic Fusarium
spp. can cause disease in both plant and animals,
including humans. The fungi are mostly present in sub-
tropical regions of the world（McGovern et al., 2001）.
The optimum temperature for infection is between 27-
31℃, but the disease can develop at a temperature that
is lower, and soil moisture levels range from 28 to 75%
with a pH of between 6-8（Glen et al., 2003）. However,
the highest damage occurs in fields where the moisture
content is low.
Fusarium wilt can remain viable in the soil for up to
30 years（Thangavelu et al., 2003）. It is destructive to
plants and progresses swiftly. By the time a plant shows
any outward sign of infection, it is already too late, and
the plant wilts and dies.
The disease and the infection behaviour of the
pathogens in planta
After conidial germination in soil, entry of the patho-
gen into the plant system is through the root tips and
is facilitated by the presence of wounds, which can be
due to nematode feeding or through vascular wounds
which can be sustained by the plant when cuttings
are procured for planting or when leaves are detached
from the stems. In the tomato-F. oxysporum f.sp.
radicis-lycopersici（FORL）pathosystem, the pathogen
multiplies abundantly through much of the root tissues,
causing severe damage including cell disorganization
through the Fusarium hyphae colonizing the roots and
cell wall alterations（M Piga et al., 1997）which may be
due to the production of plant chitinases. In addition,
during pathogen ingress in the outer root cortex there
is elaboration of callose-enriched wall appositions at
sites of attempted fungal penetration. However, pen-
etration of the callus that has grown over the wounds
caused by the fungus is impossible. Enzymes may also
facilitate Fusarium penetration into plant host（Babalola,
2010a）. It was earlier proposed that there might be
value to transforming biocontrol agents to overproduce
hydrolytic enzymes（Babalola, 2007）.
Each pathogen then grows intercellularly through the
root cortex, and, subsequently rapidly grows into the
water-conducting tissue（xylem） （Beckman, 1990）.
The microconidia produced invade the xylem of the
pseudostem where it obstructs the upward movement
of water and nutrients. Sieve cells in the stem hinder
the conidia from further spread, and, as this happens
the spores germinate and grow through the sieve cells
where they continue to germinate until the whole xylem
system is blocked. As the vessels are plugged and
collapse, the water supply to the leaves is blocked,
thus causing wilt symptoms. In most cases, the first
sign of wilt appears on one and a half month old plants
and intensifies gradually. Wilting is seen on the lower
leaves first and this later extends to the upper leaves.
Leaves, twigs or even the whole plant turns brown and
later dies and dries up（Khan and Khan, 2002）. Death
of the tomato plant is brought about by the failure of
the infected xylem to meet the water requirements of
the plant（Burgess et al., 2008）. As the plant dies, the
spores grow into the surrounding tissues where they
form chlamydospores that are released back into the
soil（Jones, 2000）. Chlamydospores can be in the
soil until there is enough moisture in the soil for them
to germinate and the process is set into motion again.
Due to the presence of antagonistic microorganisms,
insufficient moisture and alkalinity, germination can be
impaired in certain soils.
The disease or infection can be transmitted through
infected plant material and through contaminated soil.
Other means of spreading the disease is through hu-
man movement around the infected field, or the use of
irrigation water and implements previously used on an
infected crop.
Symptoms of Fusarium wilt
Plants affected with Fusarium wilt first develop stunt-
ed seedlings and yellowing of the lowest leaves that
is often restricted to one side of the plant or a single
shoot and later show the defoliation of older leaves. The
affected leaves wilt and die. Wilting progresses up the
stem until the foliage is killed and the stem decays（Lund
et al., 1998）. For this disease, brown vascular discol-
oration can be observed in stem tissue cross sections
near the soil line（Burgess et al., 2008）even though
these stems remain firm and green on the outside. In
some cases, affected plants that do not die are often
stunted and their yields are quite reduced. Where the
inoculum pressure is high, seedlings may damp off as
they emerge from the soil.
The general symptoms of many diseases of the root
and stems are yellowing, wilting and stunting while
a key symptom of pathogens which cause vascular
 INTEGRATED MANAGEMENT OF TOMATO FUSARIUM WILT
wilt disease, including Fusarium wilt pathogens, is
the browning of the internal stem（vascular）tissue
（Burgess et al., 2008）. Reddish brown discolouration
of the xylem vessels is visible inside the stem as lines
（if the stem is cut open lengthways）or dots（if it is cut
across）. On the outside of affected stems, white, pink
or orange fungal growth can be seen especially in wet
conditions.
Isolation of the pathogens from soil and infected
plants
Generally, the fungus is isolated from infected plant
tissues, which are disinfected by soaking each sample
in 1% sodium hypochlorite for 2 min. The tissues are
placed on Potato Dextrose Agar and left for incubation
at room temperature（25° C）until the growth of colonies
became visible（5 days）. Mophological identification of
F. oxysporum isolates is based on characteristics of the
macroconidia, phialids, microconidia, chlamydospores,
and colony growth traits. The forma specialis of FORL is
identified using pathogenicity tests（Hibar et al., 2007）.
Bimonthly infection and reisolation of the pathogen from
host plants is essential to maintain virulence（Babalola,
2007） .
CULTURAL, PHYSICAL AND CHEMICAL
CONTROL METHODS
According to Borrero et al.（2006）, Elmer（2006）
and L’ Haridon et al.（2011）, Fusarium wilt is a difficult
disease to control. Some of the initial management
strategies include cultural control, physical control and
chemical control.
CULTURAL CONTROL
Cultural control involves practices and farming tech-
niques that will help to increase the quality and quantity
of the yield and also reduce the influence of pests and
diseases. It involves manipulation of the environment
in non-mechanical ways to control plants pests and
diseases. It includes altering farming practices to
make the environment unfavourable for the growth of
disease pathogens and pests（Islam, 2001）. It is also
the purposeful manipulation of a garden or farm in the
growing, planting and cultivation of plants to reduce
plant disease, pest damage and numbers of pests. It
has been shown that the correct implementation of
cultural methods to control soilborne pathogens yields
improved soil structure and consequently decreases
disease incidence（Neshev, 2008）. These methods are
mainly preventive and a good knowledge of the nature,
behaviour and environmental conditions of the growth
of the disease agent is very important to controlling the
119
FIG. 1. Some of the methods used in the cultural control of
Fusarium wilt of tomato
disease development. Some of the methods used in the
cultural control of Fusarium wilt of tomato are shown in
Figure 1.
Crop rotation
Crop rotation is the practice of growing a series of
crops that are not similar in the same area season
after season. It is an easy way to control insects and
diseases at little or no cost. For example, cabbage,
tomatoes, or cauliflower planted in the same farm area
each year will actually encourage disease-causing
organisms to multiply in the soil. Plants affected by
disease should be removed and disposed of as soon
as possible, and then the soil should not be cultivated
with the affected plant type for at least 4-5 years. The
contaminated soil should be replaced with fresh soil
from another part of the garden. Disinfection of soil by
steaming or soil solarization also helps to reduce the
disease load. Crop rotation helps to reduce the amount
of pathogens in the soil. For it to effectively control a
soilborne pathogen, the pathogen must be completely
eliminated along with the plant residues from the
farmland（Neshev, 2008）
Hygiene（sanitation control）
Diseased tomato plants and plant parts, weeds
and residues should be removed and destroyed. They
should not be kept in a compost pile. Burning them
is the best, but they can also be placed inside plastic
bags and disposed as garbage or treated with a
disinfectant. Rapid crop destruction at the end of the
season is crucial for managing tomato yellow leaf curl.
Tools that have been used on the farm should always
be disinfected and cleaned before reuse. The use of
sanitized foot wear and clothing while working on the
farm helps to prevent the carrying of infected soils from
one plot of the field to another.
 120 C. AJILOGBA ET AL.
Fallowing
Fallowing is when a piece of land is left idle without
cultivation throughout the growing season. This will
reduce disease agents in the soil. Fallowing during
the summer months is helpful when soil temperatures
are high. The soil is kept dry and free of plant growth
by constantly ploughing. Exposure of the soil to high
temperature and excessive drying by ploughing helps to
destroy exposed soil-borne disease organisms, such as
nematodes. Other benefits of fallowing are insect and
weed control. Factors that encourage fallowing includes
high temperature and low rainfall.
Mulching
Mulching is the addition of a thick layer of mulch
on the soil surface to help control weed, optimise soil
moisture and keep the soil cooler than 80°F. It helps
in disease control by standing as a barrier between
the plant parts above the ground and plant pathogen
in the soil. Since it helps to control weeds, it also
helps in altering the environment for these pathogens
thereby creating unfavourable conditions for them and
controlling diseases（Raid, 2011）. In order to avoid
splashing soil borne diseases on tomato leaves during
watering, mulching of the plant is advised（Francis,
2012）. Combination of soil solarization and two layers
of mulching decrease the rate of fungal infection in
plants（Garibaldi and Gullino, 1991）.
Avoid overhead irrigation
Overhead irrigation aggravates disease problems; this
is because if the foliage is wet, fungi and bacteria are
easily spread. For example leaf spot diseases develop
rapidly when leaves are moist Overwatering should
also be avoided because excessively wet conditions
increase soil-borne fungal diseases like root rots and
wilts（Raid, 2011）. Drip drainage is recommended
（Labrada, 2008）.
Plant spacing
Plants should be properly spaced. Air circulation is
reduced with overcrowded plants and this enhances
leaf spot diseases like Septoria leaf spot on tomato.
On the other hand, with curly top virus disease,
overcrowding tomato plants reduces the disease
incidence as the insect vector finds the plants less
attractive in their crowded state. Also in a situation of
disease outbreak, overplanting of plants where possible
will ensure adequate production.
Weed control
Weeds harbor insects and serve as hosts for many
virual diseases. For most viruses to survive they must
remain in a living organism whether it be a host plant or
insect. Destroying weeds in and around the garden may
eliminate potential overwintering host plants. Destroying
susceptible diseased weeds helps to reduce the
incidence of transfer of pathogens like Fusarium spp. to
plants.
Use of disease resistant cultivars
Cultivars resistant to or tolerant to Fusarium wilt are
available for some plants, e.g. peas and China aster.
However a cultivar that is resistant to a particular forma
specialis may not be resistant to the other races of the
same forma specialis（Burgess et al., 2008）.
Excessive handling of plants
This includes tying, thinning, and pruning. Wounds
on tomato plants as a result of thinning and pruning
increase their susceptibility to Fusarium wilt. Some
diseases such as bacterial spot, tomato mosaic, and
bacterial speck can be spread while carrying out any
of these actions. Do not prune roots while cultivating
leaves because roots with cuts can create openings for
root rot and wilt fungi to enter and reduce plant vigour.
Grafting
Grafting of vegetables is available, where the
rootstock is resistant or not susceptible to Fusarium, for
example, tomatoes. Young plants, seeds and cuttings
from disease-free stocks can also be obtained from
reliable sources.
Planting on raised beds
Planting seeds on raised beds help to improve
drainage. Warmth in raised beds is higher than on level
soil and this increases the rate of emergence of the
seedlings, thus reducing their susceptibility to disease
compared to the case of slow emergence.
Choosing the best planting and harvesting times
When conditions are not favourable for the disease
or pest, crops may be planted and harvested and full
yield can still be achieved. In order to achieve this full
yield and get a head start growing, plant seeds can be
protected by using cold frames or hot caps. The crops
then have a competitive edge over the disease or pest.
In the case of pests, the farmer must have a good
knowledge of the emergence times and life cycle of
the pest to be controlled while in the case of diseases,
the farmer must have a good knowledge of the
environmental conditions that enhance the emergence
of the disease and its causal agent.
Taking advantage of alleloparthy and intercropping
Alleloparthy is a natural chemical interaction among
plants. Some plants may release toxins; decaying plant
 INTEGRATED MANAGEMENT OF TOMATO FUSARIUM WILT
tissues with the help of microorganisms can also release
toxins. In some cases, nontoxic compounds can be
modified into toxic compounds by microbial activities.
For example, the growth of winter annual weeds may be
inhibited by black walnut trees and Johnson grass and
may also offer some control of root knot nematodes.
Intercropping
Intercropping is the cultivation or planting of two or
more crops together on the same farm land or field
at the same time. It helps to maintain crop yield, soil
fertility, reduce risks, control weeds, and reduce the
incidence of pests and diseases.
For example, tomato and maize can be intercropped.
The tomato gets shade from the maize and is produced
out of season, thereby increasing the farmer s yield and
income. In Cuba this method increased annual yield by
5-6 tonnes/ha（Wolfswinkel, 2010）.
PHYSICAL CONTROL
This includes soil solarization and soil disinfection
Soil solarization
This is done by spreading a clear plastic sheet
over the soil for several weeks. This helps to trap
solar energy which in turn inhibits soilborne disease,
nematodes, insects and many weed seeds. This is
usually done during summer when the air temperature
is high and there is intense radiation（Ploeg and
Stapleton, 2001）.
Soil disinfection using heating and steam.
This is usually done as a preplanting method. Hot
water can be used according to Tetaya（2001）and
can keep the soil sterilized for up to three years. Steam
can also be used especially in greenhouse conditions
（Gullino, 2001） . Soil disinfection helps to keep the soil
sterile and free from disease causing pathogens.
CHEMICAL CONTROL
Chemical control is the use of chemicals to control
plant diseases（fungal, viral, bacterial and nematicidal）,
pest infestation and weeds. Some of these chemicals
include prochloraz, propiconazole, thiabendazole,
carbendazim, benomyl, thiophante, fuberidazole and
all of the benzimidazoles. In 2007, Nel et al reported
that benomyl was partly effcetive against F. oxysporum
f.sp cubense using the root dip treatment method.
This method was applied to using carbendazimal on
tomato seedlings infected with Fusarium wilt and it led
to about 24% increase in yield（Khan and Khan, 2002）.
Prochloraz has shown to be very effective against
121
Fusarium wilt of tomato and banana（Song et al., 2004;
Nel et al., 2007）. Fungal diseases may be controlled
through the use of fungicides and other agriculture
practices; however, new races of fungi often evolve that
are resistant to various fungicides（Sierotzki and Ulrich,
2003）. Apart from this, fungicides adversely affect
other useful soil microorganisms, affect human health
and also pollute the environment（Gerhardson, 2002）.
Chemical control measures create imbalances in the
microbial community, which may be unfavorable to the
activities of the beneficial organisms and may also lead
to the development of resistant strains of pathogens.
Pollution of the soil by overuse of chemicals has also
led to harmful effects on humans（Asaka and Shoda,
1996）.
Chemical control of the disease is not satisfactory
because generally the disease is controlled by preplant
soil fumigation with methyl bromide. In addition to
other potential health, safety and environmental risks,
methyl bromide was classified as an ozone-depleting
compound and has been banned from use（Larkin and
Fravel, 1998）. There are no effective fungicides for the
control of Fusarium wilt（Burgess et al., 2008）.
The lack of reliable chemical control agents, the
occurrence of fungicide resistance in pathogens, and
the breakdown or circumvention of host resistance
by pathogen populations are among the key factors
underlying efforts to develop other control measures
（Weller et al., 2002）. The search for alternative
strategies also has been stimulated by public concerns
about the adverse effects of soil fumigants such as
methyl bromide on the environment and human health
（Weller et al., 2002）. Since the earliest observations
of antagonist disease suppressing soil microorganisms
more than 70 years ago, plant pathologists have been
fascinated by the idea that such microorganisms could
be used as environmentally friendly biocontrol agents
both in the field and greenhouses. Thus biological
control of Fusarium wilt can offer a potential alternative
to chemical fungicides（Alwathnani et al., 2012）.
BIOLOGICAL CONTROL AND BIOLOGI-
CAL CONTROL AGENTS（BCAS）
Biological control offers a better alternative to the
use of chemicals. It is the use of natural antagonistic
organisms to combat pests or suppress plant diseases
（Lugtenberg and Kamilova, 2009）. Since PGPR（plant
growth-promoting rhizobacteria）is known for growth
promotion and disease reduction in crops, biocontrol
by use of PGPR represents a potentially attractive
alternative disease management method（Jetiyanon
and Kloepper, 2002）.
According to Jacobsen et al.（2004）, Bacillus-
 122 C. AJILOGBA ET AL.
based BCAs have great potential in integrated pest
management（IPM）systems（plant diseases included） . and direct parasitism.
Multiple Bacillus and Paenibacillus spp. can promote
crop health in a variety of ways. Some populations
suppress plant pathogens and pests by producing
antibiotic metabolites, while others may directly stimulate
nutrient uptake by plants, either promoting rhizobial and
mycorrhizal symbiosis or by fixing atmospheric nitrogen
directly（Gardener, 2004）. Specific strains of the
species B. amyloliquefaciens, B. subtilis, B. pasteurii,
B. cereus, B. pumilus, B. mycoides, and B. sphaericus
elicit significant reductions in the incidence or severity
of various diseases on a diversity of hosts. Elicitation of
Induced Systemic Resistance（ISR）by these strains
has been demonstrated in greenhouse or field trials on
tomato（Kloepper et al., 2004; Choudhary and Johri,
2009）.
Biocontrol by PGPR
PGPR can be defined as free-living rhizosphere-
occupying bacteria that enhance plant growth and can
also be classified as biocontrol agents, biofertilizers
or biopesticides depending on their activities/abilities
（Labuschagne et al., 2010）. It can also be defined
as root-colonizing bacteria（Rhizobacteria）that exert
beneficial effects on plant growth and development,
for example, in seedling emergence, colonizing roots,
stimulating overall plant growth, mineral nutrition,
and water utilization, as well as disease suppression
（Choudhary and Johri, 2009）. As they control plant
pathogens, and promote plant growth, they are also
involved with rhizomediation by degrading xenobiotic
compounds and enhancing the efficiency of fertilizers
（Kloepper et al., 2004）. Some of the reasons for the
sustained interest in biocontrol by means of PGPR
include the following:
• huge crop losses sustained due to diseases including
soilborne diseases.
• increasing production costs, especially fertilizer costs
• the global trend toward the use of more environmen-
tally friendly production methods（Labuschagne et al.,
2010） . aquaemaris, B. atrophaeus, B. boroniphilus, B. brevis, B.
Bacillus as PGPR
PGPR such as Bacillus spp. have received much
attention as biocontrol agents. They are able to produce
several broad spectrum antibiotics and have a longer
shelf life as a result of their ability to form endospores
（Cavaglieri et al., 2005）. This allows them to resist
adverse environmental conditions and permit the easy
formulation and storage of the commercial products
（Schallmey et al., 2004; Francis et al., 2010）. In
addition to their antibiotic properties, Bacillus spp.
exhibit antagonistic behaviour against fungal pathogens
by competition or exploitation, which leads to predation
Bacillus
Bacilli are Gram-positive, rod-shaped bacteria and
a member of the faranack Firmicutes. They can be
facultative anaerobes or obligate aerobes. They can
be free-living and pathogenic and test positive for
the enzyme catalase. Under stressful environmental
conditions, the cells produce oval endospores that
can remain dormant over a long period of time under
stressful and harsh environmental conditions until
favorable conditions are restored. These endospores
may remain viable for a long period of time and are
resistant to sunlight, heat and chemicals.
Origin, taxonomy and morphology of Bacillus
Ferdinand Cohn, a contemporary of Robert Koch in
1872, recognized and named the bacterium B. subtilis.
This organism is described as Gram-positive, and it
can grow in the presence of oxygen, forms endospores
in its resting phase and represents what has become
a large and diverse genus of bacteria named Bacillus,
in the family Bacillaceae. It was in 2004 that the genus
Bacillus was split into several families and genera of
endospore-forming bacteria, based on ssRNA analysis.
The taxonomic hierarchy of Bacillus according to
Bergey's Manual 2004 is kingdom: Bacteria; phylum:
Firmicutes; class: Bacilli; order: Bacillales; family:
Acyclobacillaceae（genus: Acyclobacillus）; family:
Bacillaceae（genus: Bacillus, Geobacillus）; family:
Paenibacillaceae（genus: Paenibacillus, Brevibacillus）;
family: Planococcaceae（genus: Sporosarcina）.
Bacillus distribution and environmental conditions
Bacillus spp. are widely distributed in nature, primarily
in soil（Babalola and Akindolire, 2011）, from which they
invade dust particles. Some are in water while others
are in the air. They are so numerous and include:
B. alcalophilus, B. alvei, B. aminovorans, B.
amyloliquefaciens, B. aneurinolyticus, B. anthracis, B.
caldolyticus, B. centrosporus, B. cereus, B. circulans,
B. coagulans, B. firmus, B. flavothermus, B. fusiformis,
B. globigii, B. infernus, B. larvae, B. laterosporus,
B. lentus, B. licheniformis, B. megaterium, B.
mesentericus, B. mucilaginosus, B. mycoides, B. natto,
B. pantothenticus, B. polymyxa, B. pseudoanthracis,
B. pumilus, B. schlegelii, B. sphaericus, B.
sporothermodurans, B. stearothermophilus, B. subtilis,
B. thermoglucosidasius, B. thuringiensis, B. vulgatis
and B. weihenstephanensis.
In extremely harsh conditions, B. subtilis has the
ability to form tough protective endospores. This
 INTEGRATED MANAGEMENT OF TOMATO FUSARIUM WILT
ability to withstand harsh conditions makes B. subtilis
a perfect candidate for probiotics. B. cereus is
mesophilic, and is capable of adapting to wide and
varying environmental conditions, widely distributed in
nature and commonly found in nature as a saprophyte.
It grows optimally at temperatures between 20℃ and
40℃（Vilain et al., 2006）. Since it is a soil bacterium,
it can easily enter different food plants or food made
from animals（eggs, meat, and dairy products）as they
feed on the plants. It causes about 25% of food-borne
diseases. This occurs when food has been left without
refrigeration before being served, and toxins secreted
like emetic toxins and enterotoxins into the food will
lead to food poisoning（USFDA, 2007）.
Bacillus industrial and health significance
Some Bacillus spp. are pathogenic to plants, animals,
humans and other organisms while some are very
important and useful industrially and medically.
For example, some of them secrete large amount
of enzymes that are important like the enzyme
Barnase secreted by B. amyloliquefaciens. Barnase, a
ribonuclease is important as it helps to kill plant cells
that have been infected by pathogenic fungi in other to
stop the spread of the infection. B. amyloliquefaciens
also is the natural source of amylase which is used in
starch hydrolysis. Subtilisin, a protease widely used in
commercial products, for example, in synthetic organic
chemistry research, laundry and dishwashing, skin care
ointments, detergents, food processing, cosmetics
and contact lens cleaners, can also be secreted by
B. amyloliquefaciens. Also the BamH1 restriction
enzyme used in DNA research is secreted by B.
amyloliquefaciens.
Useful antibiotics are also produced by several
TABLE 1. Some antibiotics produced by Bacillus as Biological Control Agents
Bacillus Plant pathogen Disease in plant
B. subtilis RB14 Rhizoctonia solani Damping off of
tomato
B. subtilis Fusarium oxysporum Fusarium wilt of
f.sp ciceris chickpea
B. subtilis AU195 Aspergillus flavus Aflatoxin
contamination
B. amyloliquefaciens F. oxysporum Wilt
FZB42
B. subtilis QST713 Botrytis cinerea and Damping off
R. solani
B. subtilis BBG 100 Pythium Damping off
aphanidermatum
B. cereus UW85 Phytophtora Damping off
medicagins
123
Bacillus spp like bacitracin produced by B. subtilis
and polymyxin produced by B. polymyxa. Part of the
genome of B. thuringiensis incorporated into corn and
cotton crop resulted in genetically modified plants that
are resistant to some insect pests（Bt cotton）.
Bacillus as biopesticides, biofertilizers and
bioinoculants
Biopesticides are formulations containing
microrganisms mixtures that can safely control insect
pests. For example B. thuringiensis has been formulated
into target specific and environmentally friendly products
that help to control insects and pests（Poopathi and
Abidha, 2009）. B. subtilis QST 713 has also been
formulated into biopesticides（USEPA, 2006）.
Biofertilizers are selected strains of beneficial
microbes cultured in the laboratory and packaged in
such a way that they can be applied for seed treatment
（Muraleedharan, 2010）. They are preparations
containing cells of microorganisms which may be
nitrogen fixers, phosphorus solublizers, sulphur
oxidizers or organic matter decomposers
Bioinoculants are microbial inoculants or soil
inoculants. They are microbes or beneficial endophytes
that have been prepared into microbial products to
promote plant health. Their mechanism involves the
microbes forming symbiotic relationship with the crop
or plant in question and both of them benefitting from
the association. Applied microbial inoculants help to
improve plant nutrition, and promote plant growth by
stimulating the production of plant hormone（Sullivan,
2001）.
Utilization of non-pathogenic F. oxysporum as
biological control
Metabolites produced References
Iturin A and Surfactin Asaka and Shoda,1996
Subtilin Karimi et al., 2012
Bacillomycin D Moyne et al., 2001
Bacillomycin and Koumoutsi et al., 2004
fengycin
Iturin A Kloepper et al., 2004
Mycosubtili Leclere et al., 2005
Zwittermycin A, Silo-Suh et al., 1994;
Kanosamine Milner et al., 1996
 124 C. AJILOGBA ET AL.
The vast majority of the research conducted to date
on the biological enhancement of plants with fungal
endophytes involves the use of non-pathogenic strains
of F. oxysporum. For an example, F. oxysporum Fo47
was originally isolated from disease suppressive soils
from the Chateaurenard region of France and is well-
known for its biological control properties（Fravel et
al., 2003）. Non-pathogenic strains of F. oxysporum
have the capacity to protect plants against wilt induced
by pathogenic strains of F. oxysporum. Among the
mechanisms involved in this protection, the induced
systemic resistance and priming of the plant defense
reaction could be the mode of action of Fo47（Aimé et
al., 2008）.
MECHANISMS ADOPTED BY
BIOLOGICAL CONTROL AGENTS
This is the interaction that takes place between the
pathogen and antagonist organism and also with the
crop or particular plant, and this interaction could be
direct, mixed or indirect（Pal and Gardener, 2006）.
Direct antagonism can include parasitism and predation
while indirect can include competition. Mixed path
antagonism can include amenalism（a situation when
an organism inhibits or suppresses the growth of the
other organism by producing metabolites without itself
been affected）, i.e antibiosis and lysis.
Parasitism and predation
Microbial parasitism involves the pathogen being
attacked directly or killed. This could take the form of
mycoparasitism, in which case the parasite kills the host
fungus by secreting cellwall degrading enzymes. This
could help to create holes into the host s hyphae and
cause the death of the host by absorbing its nutrients.
The death of the host can also be as a result of close
contact between the two fungi. In some instances,
the fungi penetrate, infect and damage the host plant
（Babalola, 2010a, Babalola, 2010b, Babalola, 2010c）.
Competition
As a result of the limited resources in the rhizosphere,
there is competition among the microbes for nutrients.
An example is seen with regard to the limited amount
of iron in the rhizosphere. This leads to the production
of siderosphores in some organisms to sequester
iron from the environment. When this happens, iron
is made available to the organism that produces the
siderophores while depriving the other organism; this
can lead to disease suppression of several fungal
pathogens.
Antibiosis
Antibiosis is a situation where the microbial toxins or
metabolites secreted by the antagonist organism inhibit
the growth of the pathogen or completely kill it while the
antagonist organism is not affected（Pal and Gardener,
2006）. Some antagonist organisms produce only one
metabolite while others are known to produce multiple
metabolites so they can be effective against one or
more pathogens as shown above.
Phosphate solubilization
Phosphate solubulization involves liberating soluble
phosphorus from insoluble calcium phosphate
[Ca（PO4）]. Some bacteria are able to dissolve the
mineral phosphate and make it available for plants
（Babalola, 2010d）. Solubilization of phosphate
by microbes is of practical importance in today s
agriculture. Application of the phosphate-solubilizing
microbes, Bacillus, Pseudomonas, Agrobacteria,
Aspergillus, Enterobacter and Trichoderma around the
roots of plants, in soils, and in fertilizers release soluble
phosphate, promote plant growth and protect plants
from pathogen infection.
Hydrogen cyanide production
Cyanide is a volatile inhibitor of microbial growth
（Bakker and Schippers, 1987）. HCN is a low molecular
weight metabolite, a biocide with antifungal activity
（Babalola, 2010d）. Volatiles produced by bacteria can
influence both fungal mycelial growth rate, enzyme
activity（Fiddaman and Rossall, 1995）and inhibit spore
germination in a variety of fungal species.
Induced systemic resistance
ISR refers to the ability of the plant to induce defence
responses as a result of some biotic or abiotic inducing
agent from pathogens. When plants and pathogens
interact, it could result in a response that is compatible
to both of them and lead to infection. On the other
hand it could also result into a response that is not
compatible to both organisms and lead to resistance
from the plant. The resistance can be localized or
systemic depending on the response from the plant.
When the response is incompatible, the resistance from
the plant can be an oxidative outburst that will lead
to the eventual death of the pathogen（localized）. It
could also lead to production of antimicrobial products
such as phytoalexins and pathogesis related proteins
（systemic） （Heil and Bostock, 2002）.
Kloepper et al.（2004）reported that some strains
of B. cereus, B. sphaericus, B. subtilis, B. pumilus, B.
amyloliquefaciens, B. pasteurii and B. mycoides elicited
induced systemic resistance in plants like the tomato.
Soil suppressiveness
 INTEGRATED MANAGEMENT OF TOMATO FUSARIUM WILT
Soils harbour several PGPR, some of which produce
siderophores. Suppressive soils are those in which a
specific pathogen does not persist or cause disease
despite favorable environmental conditions though
it could establish itself. In some other cases, the
disease occurs but gradually is reduced in severity
with the continuous growing of the same crop. This
suppressiveness can be inducible and durable for years.
Nonpathogenic F. oxysporum, isolated from Fusarium
wilt-suppressive soils have been effective against
Fusarium wilt of several different crops（Minuto et al.,
1995） .
CONCLUSION
Each strategy used in the control of tomato Fusarium
wilt is unique. Integrated disease management
strategies are been advocated for in such a way that
will not be harmful to the environment, useful microbes,
plants, animals and human lives.
Apart from looking at the fact that these strategies
are working in different ways, efforts should also be
on examining and developing the efficacy, efficiency
and durability of these strategies on the fields and not
just in the greenhouse. The different formulations of
microbial products that will give the most efficient result
should also be considered very important in biololgical
control while education of farmers on the appropriate
use of cultural methods and their integration into other
strategies should also be an area of interest
ACKNOWLEDGEMENTS
We are grateful to National Research Foundation,
South Africa, for a grant that supports work in our
laboratory.
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