African Crop Science Conference Proceedings, Vol. 7. pp.

1279-1283
Printed in Uganda. All rights reserved
ISSN 1023-070X/2005 $ 4.00
2005, African Crop Science Society
Occurrence and prevalence of cowpea virus diseases in uganda
M. ORAWU, R. MELIS, W. DE MILLIANO, M. LAING & E. ADIPALA
1
African Centre for Crop Improvement, University of KwaZulu Natal, Scottsville 3209, PMB, South Africa

1
Regional Universities Forum for Capacity Building in Agriculture, Makerere University, P.O. Box 7062, Kampala, Uganda
Abstract Field samples were obtained from 5-6 week old cowpea plants from 32 locations in eastern Uganda in 2004. Double
antibody sandwich enzyme-linked immunosorbet assay (DAS-ELISA) was used to test the 108 virus symptomatic leaf samples.
The virus incidences ranged from 46.6 to 96.2%, and severities ranged from 13.8 to 31.8%. The common viruses identified were
cowpea aphid-borne mosaic virus, cowpea mild mottle virus and cowpea severe mosaic virus. Of the samples tested, 21.3% were
infected by a single virus and 7.4% with two viruses. Cowpea severe mosaic virus was the most common (13% incidence) followed
by cowpea mild mottle virus (6.5% incidence) and the least was cowpea aphid-borne mosaic virus (1.9% incidence). Cowpea severe
mosaic virus occurred in mixed infections with cowpea mild mottle virus and cowpea aphid-borne mosaic virus, but mixed infection
of cowpea mild mottle virus and cowpea aphid-borne mosaic virus was rare.
Key words: Infection, occurrence, Vigna unguiculata, virus
Rsum Des chantillons taient obtenus 5-6 semaines aprs plantation du nib de 32 endroits lest de lOuganda en 2004. Une
enzyme doublement anticorps associe aux essais immunosorbet (DAS-ELISA) taient utilise pour tester le 108 me chantillon
dune feuille avec les symptme dun virus. Lincidence du virus range de 46.6 a 96.2%, et la svrit range entre 13.8 a 31.8%. Le
virus commun tait la mosaque aphide du nib, le virus du doux du nib et la mosaque svre du nib. Parmi les chantillons
tests 21.3 % taient infects par un seul virus et 7.4% avec deux virus. La mosaque du nib tait le plus commun des virus
(incidence 13%) suivi par motte douce du nib (incidence 6.5%) et la faible incidence tait celle de la mosaque aphide du nib
(1.9%). Le virus svre de la mosaque du nib apparu dans une infection mixte avec la motte douce et laphide du virus de la
mosaque. Mais une infection mixte de la motte douce du nib et la mosaque aphide du nib tait rare.
Mots cls: Infection, apparition, Vigna unguiculata, virus
Introduction
Cowpea (Vigna unguiculata L. Walp) is a major source of
protein and of considerable importance for human nutrition
in the semiarid and tropical regions of Africa (Gowda et al.,
2000). In West Africa, cowpea is second in importance after
groundnuts, with Nigeria accounting for over 70% of the
total world production (Sing et al., 2000). In Uganda,
cowpea is the third most important legume food crop after
the common beans (Phaseolus vulgaris L.) and groundnuts
(Arachis hypogea L.) (Sabiti et al., 1994). As a legume crop,
its tolerance to moisture stress and marginal soils makes it
suitable for cultivation by most farmers in Uganda. The
leaves and seeds are consumed as an important supplement
to staple cereal diet. In addition, cowpea is a major
component of the cropping system on farmers holdings
where it is grown in association with cereals such as maize,
sorghum and millet, although it is sometimes grown as a
monocrop. In eastern Uganda, where nearly 90% of the
countrys crop is produced (Adipala et al., 1997), cowpea
production is on the increase due to high demand from
both the local and external markets, and many farmers in
the region now grow cowpea for cash markets (Sabiti et al.,
1994).
However, viral diseases are among the most
agriculturally important and biologically intriguing groups
of plant pathogens, and cause serious economic losses by
reducing yields and quality of the crop (Byoung-Cheorl
et al., 2005). Cowpea yields, especially among subsistence
farmers, are generally low due to several factors, but viral
diseases remain a major constraint to production on a
large scale (Thottapilly and Rossel, 1992). Yield losses of
cowpea up to 87% due to infection by viruses have been
reported in Nigeria (Shoyinka et al., 1997), and the
complete loss of cowpea crop in northern Nigeria was
attributed to cowpea aphid-borne mosaic virus disease
(Raheja and Leleji, 1974).
Worldwide, up to 20 viruses have been reported to
occur on cowpea, but only eight viruses are known to
infect cowpea in Africa (Thottappilly and Rossel, 1992).
The economically important viruses in Africa include
cowpea aphid-borne mosaic virus potyvirus, cowpea mild
mottle virus carlavirus, cowpea mosaic virus comovirus,
cowpea chlorotic mottle virus carmovirus, cowpea
golden mosaic virus germinivirus, southern bean mosaic
virus sobemovirus, cucumber mosaic virus cucumovirus
and tobacco mosaic virus tobamovirus (Thottappilly and
Rossel, 1992; Alegbejo and Kashina, 2001).
In Uganda, studies by Edema et al. (1997) showed
cowpea aphid-borne mosaic virus as the most common
virus, yet little attention has been given to it. Information
on the occurrence of other viruses in the cowpea growing
regions in Uganda is not well known. With changes in
the cropping systems and population dynamics of virus
M. ORAWU et al.
1280
vectors may influence virus occurrence (Wisler et al.,
1998), and hence cowpea viruses have increasingly
become very significant in Uganda. In order to provide
information that would be essential for determining
priorities in breeding for resistance to viruses, attempts
were made to verify and identify the occurrence of the
economically important viruses on cowpea in eastern
Uganda.
Materials and methods
Field survey and sampling. Field surveys were conducted
on farmers fields during the growing season in October
and November, in the eastern Uganda. In 2004, surveys
were restricted to Soroti, Kumi, Pallisa and Tororo, the
major cowpea producing districts, where 32 locations were
surveyed. Cowpea fields in each district were surveyed
and sampled at approximately 5 km apart. About five to six
weeks old cowpea trifoliate leaves were sampled in the
four districts and a diagonal pattern of sampling was used.
The total number of plants within a quadrant (1 x 1 m) of
each field was counted approximately 30 plants and virus-
like symptomatic plants recorded. Thus, 120 plants were
recorded in each field of four quadrants. The virus
incidence was calculated as percentages of the diseased
plants over the total number of plants assessed as the
average of the cowpea fields per district. Disease severity
was also assessed visually during the study as the
proportion of the leaves with virus symptoms on
percentage score of 0, 5, 20, 25, 30, 40, 45, 50, and 60.
Serological tests. One hundred eight virus symptomatic
leaf samples were collected from 32 locations in four
districts namely Soroti, Kumi, Pallisa and Tororo. Fully
expanded trifoliate leaves from each quadrant were
sampled in a field of four quadrants and the samples were
placed separately in small plastic polythene bag. The
samples were then taken to Makerere University Biotech
laboratory and frozen at -20
0
C before being subjected to
DAS-ELISA. Double antibody sandwich enzyme-linked
immunosorbet assay was used for serological testing of
plant samples and was carried out using kits provided by
DSMZ in Germany. The kits included specifically to
particular virus antiserum namely cowpea aphid-borne
mosaic virus (CABMV), cowpea chlorotic mosaic virus
(CCMV), cowpea mild mottle virus (CMMV), cowpea
mosaic virus (CMV) and cowpea severe mosaic virus
(CSMV). Three leaf-discs were taken from top, middle and
lower parts of the leaf preferably with virus symptoms.
The kits contained positive and negative controls of dried-
ground leaf samples. The positive and negative reactions
were visually assessed, the degree of yellow coloration
determining those regarded as positive with the ELISA
test. The absorbencies were recorded at 405nm (A
405
) after
90 minutes substrate incubation period using a microplate
reader. Readings indicating twice the values of the negative
controls were considered positive.
Results
Virus disease occurrence. Figure 1 shows the mean
incidence and severity of virus symptoms that were
observed in the surveyed districts of Soroti, Kumi, Pallisa
and Tororo though there were considerable differences in
occurrence and prevalence among the districts. The
highest disease incidence was 96.2% with severity of
31.8% in Pallisa and lowest disease incidence levels of
46.6% with severity of 28.8% being recorded in Kumi (Fig.
1). However, Tororo district that recorded high disease
incidence levels of 85.8% had the lowest severity levels
of 13.8% compared to Soroti, Kumi and Pallisa districts.
Virus detection. One hundred eight leaf samples collected
from four cowpea growing districts were subjected to
DAS-ELISA tests using antisera to CABMV, CCMV,
CMMV, CMV and CSMV. Thirty five samples of the total
108 collected were virus infected. Three viruses namely
CMMV, CABMV and CSMV were detected in the samples.
Cowpea severe mosaic virus (CSMV) was detected in the
samples in the four districts surveyed. A total of 24 (22.2%)
0
20
40
60
80
100
120
Soroti Kumi Pallisa Tororo
D
i
s
e
a
s
e

i
n
c
i
d
e
n
c
e

(
%
)

a
n
d

s
e
v
e
r
i
t
y

(
%
)
Disease incidence (%)
Disease severity (%)
Figure 1. Percentage incidence and severity of cowpea plants with viruslike symptoms in the major cowpea growing districts in eastern Uganda.
1281
Occurrence and prevalence of cowpea virus diseases
symptomatic samples reacted positive with CSMV
antibodies thus making CSMV the most prevalent virus in
the districts (Table 1). This was followed by CMMV with
a total of 7 (6.5%) symptomatic samples occurring only in
three districts, with the exception of Tororo district.
Cowpea aphid-borne mosaic virus was detected in 4 (3.7%)
diseased plant samples obtained from Pallisa and Tororo
districts, and it was the least frequent virus among the
viruses detected.
Single and multiple virus infections. A proportion of
21.3% of symptomatic samples were infected by a single
virus, whereas 7.4% were infected with two viruses. The
most common with single infection was that of cowpea
severe mosaic virus (13%), followed by cowpea mild mottle
virus (6.5%) and the least was cowpea aphid-borne mosaic
virus (1.9%). However, cowpea severe mosaic virus
occurred in mixed infections with other viruses namely
cowpea severe mosaic virus and cowpea mild mottle virus
(5.6%), and cowpea severe mosaic virus and cowpea
aphid-borne mosaic virus (1.9%) (Fig. 2).
Discussion
Cowpea viruses have become increasingly important in
the cowpea growing regions in Uganda and subsequently
inflicted heavy losses to yields. The results reported herein
indicate substantial incidence and severity of viruses
during the survey conducted in the cowpea growing
districts in eastern Uganda. There were considerable
variations of incidences and severities among the districts
surveyed. For instance, virus incidences ranged between
46.6% and 96.2%, and severities ranged between 13.8%
and 31.8%. The incidences were high in Pallisa and Tororo
districts compared to Kumi and Soroti districts. These
variations in occurrence under different agro-ecological
environments can be due to the fact that the low virus
incidences in Kumi and Soroti may suggest presence of
low virus vectors and consequently low virus inoculum
source to cause high incidence in cowpea grown fields.
Elsewhere, studies have shown that with changes in the
farming systems, cultivar types and dynamic population
of virus vectors may affect virus occurrence (Wilser et al.,
1998). Edema et al. (1997) and Shoyinka et al. (1997)
reported the effect of weather conditions within seasons
and cropping systems on distribution of viruses in the
different environments. In other studies, however,
manipulation of plant populations especially at relatively
high plant densities also showed to affect the dynamics
of virus vectors that transmit viruses to plant crops
(Ogenga-Latigo et al., 1992a,b). Similarly, according to
Mukankusi et al. (1999) on groundnut rosette disease and
Orawu et al. (2001) on cowpea viral diseases, also reported
low virus incidences under high plant populations,
suggesting that unfavourable microclimate disfavours
virus vectors to colonise plants.
Three cowpea viruses namely cowpea aphid-borne
mosaic virus, cowpea severe mosaic virus and cowpea
Table 1. Number, percentage incidence and virus types when serologically tested in the symptomatic samples collected in four districts of Uganda.
District Samples tested Virus serological detection by DAS-ELISA
CABMV CCMV CMMV CMV CSMV
Soroti 32 - - 1 (3.1) - 11 (34.4)
Kumi 32 - - 4 (12.5) - 1 (3.1)
Pallisa 22 2 (9.1)* - 2 (9.1) - 7 (31.8)
Tororo 22 2 (9.1) - - - 5 (22.7)
Total 108 4 0 7 0 24
* Figures in parentheses are percentage incidence. - Indicate no virus was detected in the samples in any of the districts.
0
2
4
6
8
10
12
14
CSMV CMMV CABMV CSMV +CMMV CSMV +CABMV
P
e
r
c
e
n
t
a
g
e

p
l
a
n
t
s

i
n
f
e
c
t
e
d
Figure 2. Occurrence of single and multiple virus infections in symptomatic cowpea plants.
M. ORAWU et al.
1282
mild mottle virus were detected in the survey of
symptomatic cowpea plants, suggesting their existence
in the major cowpea growing areas in eastern Uganda.
Cowpea severe mosaic virus was the most common virus
while cowpea mild mottle virus was the second most
prevalent virus detected in all surveyed cowpea growing
districts. Our results suggest that cowpea aphid-borne
mosaic virus was the third and least frequent virus
detected in the samples from the four surveyed districts.
Although an earlier report by Edema et al. (1997) indicated
cowpea aphid-borne mosaic virus as the most common
virus in Uganda, but on contrary our results showed
cowpea severe mosaic virus as the common virus based
on four surveyed districts in eastern Uganda. From the
surveyed districts, antibodies of cowpea chlorotic mosaic
virus and cowpea mosaic virus were included in the
serological tests of cowpea samples, but no samples
reacted with antisera of these viruses, suggesting that
they may not be present in Uganda.
Multiple-virus infections are common among samples
from field grown cowpeas and are known to modify and
complicate symptoms, thus precluding in field diagnosis
based on symptoms (Shoyinka et al., 1997). In the study,
however, mixtures of viruses were observed in only two
of the 108 cowpea samples. The shared viruses in the
samples were cowpea severe mosaic virus and cowpea
mild mottle virus, and cowpea severe mosaic virus and
cowpea aphid-borne mosaic virus. These results compare
well with other studies on virus occurrence in mixed
infections (Shoyinka et al., 1997). The higher incidence of
cowpea severe mosaic virus (13%) in the samples
compared to cowpea mild mottle virus (6.5%) and cowpea
aphid-borne mosaic virus (1.9%), could suggest its relative
abundance of beetle vectors to survive under adverse
environmental conditions over the white fly and aphid
vectors in the surveyed districts. However, there was no
association between cowpea mild mottle virus and cowpea
aphid-borne mosaic virus in cowpea samples, probably
the aphid vectors and white fly vectors are not linked
together to cause synergism effect in cowpea plants.
Generally, the study confirmed three economically
important cowpea viruses and their relative complexes in
the cowpea fields. This situation demands that urgent
attention should be given to strengthening the
management of cowpea viruses in Uganda. The only
intervention to enhance production could be through the
use of resistant varieties, but this is often compounded
by pathogenic variability and occurrence of virus mixtures.
However, exploitation of cowpea genotypes with multiple
resistance genes would offer a better option in the breeding
work to control several viruses so as to circumvent the
escalating problems of viruses and ultimately yield losses.
Acknowledgements
This study was funded by the Rockefeller Foundation
through the African Centre for Crop Improvement in South
Africa. We gratefully appreciate the assistance extended
by director of SAARI and the Makerere University,
Department of Crop Science for the facilities provided for
the work. We also thank the support of the farmers who
allowed us free access to their fields to conduct the
surveys.
References
Adipala, E., Obuo, J .E. & Osiru, D.S.O. 1997. A survey of
cowpea cropping systems in some districts of Uganda.
African Crop Science Conference Proceedings 3, 665-
672.
Alegbejo, M.D. & Kashina, B.D. 2001. Status of legume
viruses in Nigeria. J ournal of Sustainable Agriculture
18, 55-69.
Byoung-Cheorl, K., Inhwa, Y. & Molly, M.J. 2005. Genetics
of plant virus resistance. Annual Review of
Phytopathology 43, 581-621.
Edema, R., Adipala, E. & Florini, D.A. 1997. Influence of
season and cropping system on occurrence of cowpea
diseases in Uganda. Plant Disease 81, 465-468.
Gowda, B.S., Miller., J.L., Rubin, S.S., Sharma, D.L. & Timko,
M.P. 2000. Isolation, sequencing and mapping of
resistance gene analogs from cowpea (Vigna
unguiculata L. Walp). In: C.A. Fatokun, S.A.Tarawali,
B.B. Singh, P.M. Kormawa and M. Tawo (eds).
Challenges and opportunities for enhancing
sustainable cowpea production. Proceedings of the
World Cowpea Conference III held at the International
Institutes of Tropical Agriculture Ibadan, Nigeria, 4-8
September 2000. pp. 167-184.
Mukankusi, C., Adipala, E., Kyamanywa, S., Epieru, G.,
Odeke, V., Warren, H.L. & Wilson, H.R. 1999. Effect of
host genotype, time of planting and spacing on
epidemics of groundnut rosette and Cercospora leaf
spot diseases in eastern Uganda. African J ournal of
Plant Protection 9, 37-53.
Ogenga-Latigo, M.W., Ampofo, J.K.O. & Baliddawa, C.W.
1992a. Influence of maize row spacing on infestation
and damage of intercropped beans. I. Incidence of
aphids. Field Crops Research 30, 111-121.
Ogenga-Latigo, M.W., Baliddawa, C.W. & Ampofo, J.K.O
1992b. Influence of maize row spacing on infestation
and damage of intercropped beans by the bean aphid
(Aphis fabae Scop.). II. Reduction on bean yields. Field
Crops Research 30, 123-130.
Orawu, M., Adipala, E. & Warren, H. 2001. Effect of spacing
and time of planting on occurrence of cowpea diseases
in eastern Uganda. African Crop Science Conference
Proceedings 5, 399-407.
Raheja, A.K. & Leleji, O.I. 1974. An aphid borne mosaic
disease of irrigated cowpeas in northern Nigeria. Plant
Disease Reporter 58, 1080-1084.
Sabiti, A.G., Nsubuga, E.N.B., Adipala, E. & Ngambeki,
D.S. 1994. Socio-economic aspects of cowpea
production in Uganda: A Rapid Rural Appraisal.
Uganda J ournal of Agricultural Sciences 2, 29-35.
Shoyinka, S.A., Thottappilly, G., Adebayo, G.G.& Anno-
Nyako, F.O. 1997. Survey on cowpea virus incidence
and distribution in Nigeria. International J ournal of
Pest Management 43, 127-132.
1283
Occurrence and prevalence of cowpea virus diseases
Singh, B.B., Ehlers, J .D., Sharma, B. & Freire Filho, F.R.
2000. Recent progress in cowpea breeding. In: C.A.
Fatokun, S.A.Tarawali, B.B. Singh, P.M. Kormawa and
M. Tawo (Eds). Challenges and opportunities for
enhancing sustainable cowpea production.
Proceedings of the World Cowpea Conference III held
at the International Institutes of Tropical Agriculture
Ibadan, Nigeria, 4-8 September 2000. pp. 22-40.
Thottappilly, G. & Rossel , H.W. 1992. Virus diseases of
cowpea in tropical Africa. Tropical Pest Management
38, 337-348.
Wisler, G.C., Duffus, J.E., Liu, H-Y. & Li, R.H. 1998. Ecology
and epidemiology of whitefly-transmitted
closteroviruses. Plant Disease 82, 270-280.