|Research Article (Open access)
SSR Inst. Int. J. Life Sci., 8(6):
First Record of A
serious Wilt Disease Induced by Fusarium sacchari and Root Borer on
Sugarcane in Kenana Scheme, Sudan
Elnair Akasha Mohammed Akasha1*, Elnaeem
Mohamed Abdelrahim1, Ahmed Hashim Ahmed2
1Department of Crop Protection, Research and Development
Department, Kenana Sugar Company, Sudan
2Department of Crop
Protection, Faculty of Agricultural, University of Khartoum, Khartoum, Sudan
Akasha Mohammed Akasha, Department of Crop Protection, Kenana Sugar Research
and Development, Kenana, Sudan
ABSTRACT- Background: Wilt of sugarcane was recorded almost 100 years ago and is one of
the major fungal diseases affecting cane production and productivity. Many
commercial varieties were withdrawn from cultivation due to their
susceptibility to the disease in many countries. Even though the disease was
recorded long back.
Methods: For the assessment of disease status and
varieties susceptibility of wilt disease, an extensive survey was conducted on
sugarcane fields of 2020-2021 of Kenana Sugarcane Company. Isolation of the Pathogen
was done by two types of culture in Petri dishes containing PDA the other was
incubated in a Plotter test. Both segments were incubated at 28±2°. The
pathogenicity test was conducted according to Koch's postulate by Dipping
inoculation method and the Plug inoculation method.
Results: The result of the survey revealed that
the symptoms of the disease appeared clearly in the fields and the result of
laboratory isolation indicated that the pure culture of causing pathogen, Fusarium
sacchari based on detailed culture morphology. Among the insect pests, root
borer was found to associate with wilt disease from sugarcane-infected stalks.
The result of the pathogenicity test in pot culture under greenhouse conditions
using the variety Co 997 with the pathogenic culture of F. saccharri
induced the same symptoms. And re-isolation of the pathogen is similar to the
isolation from fields.
Conclusion: We were concluded that the wilt of
sugarcane affects cane productivity in Sudan and this is considered as the
first record of wilt disease in sugarcane in Sudan. As sugarcane is an
important crop, future disease control methods will soon be started to control
this important disease.
INTRODUCTION- Sugarcane (Saccharum
officinarum) is grown in the tropics and subtropical regions of the world.
It was estimated that sugarcane was cultivated on over 26 million hectares in
more than 90 countries with a worldwide harvest of 1.84 billion tons. Sugarcane
cultivation is challenged by several biotic and biotic factor. In addition to
other abiotic factors like draught, unseasonal floods etc sugarcane suffers
from several diseases caused by fungi, bacteria, viruses, nematodes, and
of sugarcane caused by Fusarium moniliforme var. sub glutinans cause
serious quantitative and qualitative losses which have negative effects on
sugarcane production, as well as in the sugar industries . The
most important diseases in Kenana Sugar Company are smut and ratoon stunting
disease . For the first time described a stem rot disease in
India in sugarcane under the term wilt and noted Cephalosporium sacchari
as the causal agent. However, recorded stem rot of the basal portion of
unwounded sugarcane stem having species of Fusarium is associated with disease
in Barbados . An overall 20.22% loss of cane is caused by
different disease, which worked out to Tk. 250 corers annually. 
Fungal diseases are one of the major concerns to agricultural production. Out
of 40 sugarcane diseases in Bangladesh, wilt is considered as one of the most
damaging . Although there
were report on the disease incidence  made detailed surveys in
all the sugarcane growing areas and they reported that the disease severity is
highest in Gujarat state, east coastal regions in Tamil Nadu, Andhra Pradesh,
and Orissa in tropical India. No definite role of nematodes in stimulating the
wilt syndrome in sugarcane is established. However, Viswanathan et al. 
reported a possible association between nematode and wilt fungi. They recorded
highest (65%) wilt incidence and reduction in fresh plant weight when sugarcane
plants were inoculated with nematodes and wilt fungi (Fusarium sacchari
and Acremonium implicatum) together.
Detailed studies have been carried out on phenotypic and genotypic
variability at this Institute . Phonotypical characterization of
the pathogen was done based on growth rate, pigmentation, texture, nature of
phialides and conidia produced. Cultural characteristics of 117 isolates have
been studied and they were grouped in to three groups based on the growth rate.
Based on the mycelium colour, the isolates were categorized into 7 group’s
viz., white, orange, orange-pink, pink, dark pink, pinkish violet and reddish
brown. More than 75% of the isolates showed typical pinkish pigmentation and
other cultures exhibited varying shades of pinkish pigmentation .
Wilt fungus in association with some insect pests of sugarcane, particularly
stalk borer and scale insects, causes significant damage to the crop. In
association with stalk borer, the disease has been reported to bring a loss of
about 8.75 tons/ha . Although the disease affects the crop
during different phases viz., germination, young crop and maturity, disease
expression/symptom are well known in mature crops as yellowing followed by
drying of foliage and subsequent withering of infected plants. The germinating
setts or germinated settlings also show the disease symptoms and usually, such
cases are ignored . Apart from separate infections of F.
sacchari, combined infections of F. sacchari and Colletotrichum
falcatum causing red rot is found under field conditions in different
states in India  or Ceratocystis paradoxa cause much more
damage than when present alone . Found synergistic effect of
combined inoculation in disease susceptible clones. Fusarium produced a mild
infection, which did not extend beyond the inoculated internode even after
three months. In continuation with C. falcatum, however, it led to
severe infection in all the varieties and the advance of the lesion is greater
in the combined infection than infection by C. falcatum alone. Between
the two pathogens, red rot caused greater deterioration in the quality of
sugarcane than wilt . The maximum inhibition zones of the
pathogen causing wilt and root rot disease of sugarcane were recorded by
Acetobacter, PSB and Trichoderma. Similar results were recorded by Sabalpara
et al.  noticed that
for the purpose of controlling sugarcane wilt and root rot pathogen and to
obtained efficient isolates, of Trichoderma were tested. The results indicate
that all the tested isolates were effective.  Reported that the
study showed that the different Trichoderma isolates have good antagonistic
effect on the mycelium growth of Fusarium sp. and Pythium sp.
These results are in accordance with earlier investigation. The objective of
this manuscript is to investigate the presence of wilt disease in sugarcane and
MATERIALS AND METHODS- To assess the disease status
and varieties susceptibility of wilt disease, an extensive survey was conducted
on sugarcane fields 2020–2021 of Kenana Sugarcane Company located between
latitudes 13:10–12.350 North and longitudes 32:40–32:550
East, it is about 410 meters above sea level, within the tropic semi/arid
climatic zone. A photo was taken from effected stalk also random sample was
taken for further investigation. During the survey.
Isolation of the Pathogen- The infected stalks cut into 3
to 4 cm long were surface sterilized by sodium hypochlorite solution 4% for 1
to 2 minutes, washed four times with sterilized distilled water to remove sodium
hypochlorite solution and left to dry. Then the sample was divided into two subsamples.
Six segments from the subsample were incubated in each Petri dish containing
PDA. Similar segments were incubated in the Plotter test. Both segments were
incubated at 28±2°C temperature and inspected daily for fungi growth.
Pathogenicity Test- Koch's
postulates, firstly the cultivars used in the pathogenicity test should be
identical to those on which the disease has been observed and isolated from the
field. Then, when the cultures were inoculated into susceptible plants, they
must initiate the characteristic disease symptoms. Finally, the organisms were
re-isolated in pure culture and re-identified, after which it must be similar
to the original organism that had been observed  before each step
is followed correctly and if produced the identical pathogen after re-isolation,
then the pathogenicity test had been succeeded  the
pathogenicity test was done by two methods described as the following:
Dipping inoculation method- Twenty
setts from a nursery of sugarcane variety Co 997 was catted as single bud
washed by running tab water and then divided into two group ten setts dipped in
spore’s suspension concentrated 3x105 for ten mints and ten setts
dipped in sterile distilled water for ten mints and then planted in a plastic
bug in the greenhouse for further investigation. Observation of the disease
development was started after one month up to six months of inoculation and at six-month,
inoculation stalks of the cane were cut and split open to investigate the
internal symptoms .
Plug inoculation method- In the plug
method, the inoculum was placed in the bore hole and sealed with modelling parafilm
to avoid aerial contamination ten cane (stalks) were inoculated in the
internode number three from the base of the stalks beside the land, spores’
suspension as concentration 3x105, about 1 ml of conidial suspension
was placed in the node. Ten stalks as control were treated with sterile
distilled water. Observation of the disease development was started after two weeks
up to six months of inoculation, inoculation stalks from treated and untreated
cane were cut and split open for final observation .
RESULTS- The field surveys were conducted in Kenana Sugar Company (KSC) to
investigate the occurrence of wilt disease in sugarcane. The wilt stalks were
observed dry from the underside part of the stalks and spread upward, when
splitting stalks longitudinally the internal symptoms were pith tissue, light
to dark purplish-brown. Varying shades of pinkish-red or brownish-red
discolouration were seen in the internodes. Stunted with yellowing and
withering of crown leaves. The midribs of all leaves in a crown generally turn
yellow, while the leaf lamina remains green. The leaves dry up and stalks
develop hollowness in the core. The core shows reddish discolouration with
longitudinal red streaks passing from one internode to another. In severe
cases, spindle-shaped cavities tapering towards the nodes develop in each
internode. The canes emit a disagreeable, with a lot of mycelia threads of the
fungus covering the cavity (Fig. 1). The culture appears in pure hairy white mycelium
(Fig. 2), and the pathogen was identified as F. sacchari based on
microscopic features. Morphological features of the fungus: the mycelium of the
fungus was white cottony, with culture pigmentation peach salmon, vinaceous
purple violet, macroconidia general morphology relatively slender, slightly
falcate and thin-walled, and number of septa: Usually 3-4 septate. Macroconidia shape and septation: oval, slender,
and primarily 0-septate, although 1- or 2-septate elongated. Chlamydospores, absent
Fig. 3 and also the result of the survey indicated that the wilt disease is
associated with root borer as shown in Fig. 4.
Fig. 1: Infected
Stalks of Sugarcane by Wilt Disease
Fig. 2: White Hairy Mycelium of Fusarium sacchari
Fig. 3: Macro and Microconidia of Fusarium sacchari
Fig. 4: Root borer associated with wilt disease on
Pathogenicity test- The pathogenicity test was conducted according to Koch's
postulates, where the steps are shown in Fig. 5 as followed:
Step A shows the infected stalks collected
from sugarcane fields in Kenana State showed symptoms of wilt disease.
showed that the fungi isolated from infected grass in step A grew in culture.
Step C infection
of Fusarium sp, the result shows typical symptoms as shown in step A.
Step D the results of re-isolation of the
fungi from inoculated healthy stalks in step C the pathogens showed the typical
characteristic of the isolated fungi from infected sugarcane stalks (the spores
and mycelia of species) as shown in the following.
Fig. 5: Pathogenicity Test Cycle According to Koch's Postulates
Common Topics during Health Talks/ Hospital Visit- The
findings show that cervical or breast cancer screening was never a topic for
discussion. Some of the participants stated: “We are told to take care of body.
We are told not to be lazy. Eat good food such as egg, vegetables, fish, liver,
and so on”.
Awareness of Cancer, majority of the participants did not hear about cervical
cancer, only a few admitted to have heard about Breast Cancer. Some of the
participants gave responses like this: “We don’t know about it; I have not
heard about cancer of cervix; I only heard of Breast cancer; I am hearing it
for the first time”. Only a few women said that: “I heard it destroys the mouth
of the womb and the person will not be able to deliver a baby and will
eventually lead to the evacuation of the womb”.
The survey of commercial sugarcane fields in KSC
revealed that the wilt disease of sugarcane was present in the variety Co 997 according
to the result of the plant's pathology laboratory and confirmed by
pathogenicity test, the main causes of this disease according to the test is F. sacchari. Also, some insect pest was
found associated with wilt disease as root borer. Butler and Khan  studied
wilt in detail and described Cephalosporium sacchari as the associated
pathogen. Subsequently, several workers reported Fusarium moniliforme var sub-glutinans as the causative pathogen. 
Coined a new species F. sacchari. To
which both C. sacchari and F. moniliforme var. sub-glutinans were
made synonyms. Later  distinguished two varieties of F. sacchari namely, F. sacchari var. sacchari and F.
sacchari var. sub-glutinans, the former having mostly septate conidia in
the aerial mycelium and without sporodochia, while the latter with 1-3 septate
conidia, macroconidia more commonly formed in sporodochia. Similar to many
vascular wilts in other crops, the wilt of sugarcane is a major disease
affecting cane productivity in India. The survey for wilt incidence across the
country revealed that the pathogen is widespread in the country [18-20].
Although the disease was reported a century ago. Available information on the associated
pathogen is scanty, shallow and contradictory; hence a detailed investigation
was carried out to characterize the pathogenic isolates by cultural,
morphological and molecular methods after a comprehensive survey of the disease
in major sugarcane growing states. The pathogen exhibited enormous variation in
cultural characters and that could not be used to characterize the isolates.
Using morphological features and molecular profiles in ISSR and IGSRFLP, the
pathogen was identified as F. sacchari.
Wilt fungus in association with some
insect pests of sugarcane, particularly stalk borer and scale insects, causes
significant damage to the crop. In association with stalk borer, the disease
has been reported to bring a loss of about 8.75 tonnes/ Viswanathan 
and Viswanathan and Rao  reported that the weight decline was
24.9% when the mean incidence of stalk borer-wilt complex was 51.4% .
Reported a high incidence of wilt (90%) in association with stalk borer in the
cv Co 1148 and the crop was almost unfit for milling. With conservative
estimates of a loss of 3-6 tonnes/ha, the disease may cause an annual loss of
12.7-25.40 million tonnes in different years. Hence, the loss to sugarcane
production would be between Rs. 1250-2500 crores per annum .
The loss in production is borne by both the farmers and the sugar
industry. The impact of wilt-infected canes on recoverable sugar in the mill is
not assessed properly and sugar mills experience these unaccounted losses every
year. In recent years, the sudden outbreak of pokkah boeng across the country
was noticed in several varieties . It was found that Fusarium sp. associated
with pokkah boeng also caused stalk infections and produced wilt in certain
varieties. It is expected that further studies in this area would bring a new
dimension to the Fusaria associated with pokkah boeng and wilt. Eldana
saccharina is associated with Fusarium species in maize  and
sugarcane . In maize, endophytic colonization by F.
verticilliodes was correlated with higher E. saccharina infestation
and damage compared with plants treated with fungicide, suggesting a beneficial
relationship between the fungus and the insect . In sugarcane, E.
saccharina damage is associated with infection by Fusarium sp. where
larval borings facilitate access of the fungus to the inner stalk resulting in
the rot of tissue surrounding the insect borings .
CONCLUSIONS- Wilt disease symptoms in sugarcane are very common in the variety
Co 997 in Kenana Sugar Company-Sudan. Several investigations revealed the
occurrence of wilt disease induced by F. sacchari was responsible for
wilting of sugarcane stalks and this disease was associated with root borer mix
infection leading to dryness of the whole stalk. Similar to many vascular wilts
in other crops, the wilt of sugarcane is a major disease affecting cane
productivity in Sudan and this is considered the first record of wilt disease
in sugarcane in Sudan.
The survey for wilt disease in Kenana Sugar Company revealed that the
stalks rot of basal portion of unwounded stalks. As sugarcane is an important cash crop in the
Sudan, future disease control methods will soon be started to control this
CONTRIBUTION OF AUTHORS
Research concept- Elnair Akasha Mohammed
Research design- Elnair Akasha Mohammed
Supervision- Prof.Ahmed Hashim Ahmed
Materials- Elnair Akasha Mohammed, Elnaeem Mohamed
Data collection- Hawa Ahmed Ibrahim, Abu Bakr Abdalla Ali, Elnaeem Mohamed and Elnair
Data analysis and
interpretation- Elnair Akasha Mohammed
Literature search- Elnair Akasha Mohammed
Writing article- Elnair Akasha Mohammed
Critical review- Prof.Ahmed Hashim Ahmed
Article editing- Prof.Ahmed Hashim Ahmed
Final approval- Prof.Ahmed Hashim Ahmed
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