Vous êtes sur la page 1sur 5

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

Résumé Des échantillons étaient obtenus 5-6 semaines après plantation du niébé de 32 endroits à l’est de l’Ouganda en 2004. Une enzyme doublement anticorps associée aux essais immunosorbet (DAS-ELISA) étaient utilisée pour tester le 108 ème échantillon d’une feuille avec les symptôme d’un virus. L’incidence du virus rangée de 46.6 a 96.2%, et la sévérité rangée entre 13.8 a 31.8%. Le virus commun était la mosaïque aphide du niébé, le virus du doux du niébé et la mosaïque sévère du niébé. Parmi les échantillons testés 21.3 % étaient infectés par un seul virus et 7.4% avec deux virus. La mosaïque du niébé était le plus commun des virus (incidence 13%) suivi par motte douce du niébé (incidence 6.5%) et la faible incidence était celle de la mosaïque aphide du niébé (1.9%). Le virus sévère de la mosaïque du niébé apparu dans une infection mixte avec la motte douce et l’aphide du virus de la mosaïque. Mais une infection mixte de la motte douce du niébé et la mosaïque aphide du niébé était rare.

Mots clés: 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 country’s 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

1280

M. ORAWU et al.

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%)

Disease incidence (%)Disease severity (%)

Disease severity (%)Disease incidence (%)

120 100 80 60 40 20 0 Soroti Kumi Pallisa Tororo Disease incidence (%) and
120
100
80
60
40
20
0
Soroti
Kumi
Pallisa
Tororo
Disease incidence (%) and severity (%)

Figure 1. Percentage incidence and severity of cowpea plants with viruslike symptoms in the major cowpea growing districts in eastern Uganda.

Occurrence and prevalence of cowpea virus diseases

1281

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.

14 12 10 8 6 4 2 0 CSMV CMMV CABMV CSMV + CMMV CSMV+
14
12
10
8
6
4
2
0
CSMV
CMMV
CABMV
CSMV + CMMV
CSMV+ CABMV
Percentage plants infected

Figure 2. Occurrence of single and multiple virus infections in symptomatic cowpea plants.

1282

M. ORAWU et al.

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. Journal 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 Journal 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 Journal 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 Journal of Pest Management 43, 127-132.

Occurrence and prevalence of cowpea virus diseases

1283

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.