Field Studies of Cross Protection with Cassava Mosaic Geminiviruses in Uganda

The effect of cassava mosaic virus disease (CMD) was compared on plants grown from cuttings that were initially virus‐free at planting and those infected with a mild strain of East African cassava mosaic virus‐Uganda (EACMV‐UG). All initially healthy plants developed CMD symptoms within 5 months of planting (MAP) at both trial sites in Uganda, although spread was more rapid at Kamuli than at Serere. Significantly (P < 0.001) higher symptom severity scores were recorded in initially healthy plants, which had average scores of 3.6 and 3.5 at Kamuli and Serere, respectively, compared with 2.8 for mildly diseased plants at each location. Severity scores of 4 and 5 were more frequent in initially healthy plants, accounting for 77 and 39% of the total infections recorded in comparison with 47 and 11% in mildly diseased plants at Kamuli and Serere, respectively. Mildly diseased plants were significantly taller than initially healthy plants 8 and 12 MAP at both locations. However, the converse was true 4 MAP although differences were significant at Serere but not at Kamuli. Mildly diseased plants yielded significantly more tuberous roots than initially healthy plants at Kamuli but not at Serere. Average total weights of tuberous roots per plant were 2.48 and 1.63 kg for mildly diseased and initially healthy plants at Kamuli and 4.46 and 4.61 kg at Serere, respectively. These results may help to explain the increased prevalence in recent years of mildly diseased plants of local CMD‐susceptible cultivars in eastern Uganda, from where these varieties virtually disappeared following the severe CMD epidemic in the 1990s. The results also provide the first field evidence of a cross protective effect of mild strains of a cassava mosaic geminivirus.     

Reaction of Cassava Genotypes to the Cassava Mosaic Disease in Three Distinct Agroecologies in Nigeria

Nine cassava genotypes were grown at six representative sites in Nigeria for 3 years to study their response to cassava mosaic disease (CMD), investigate the influence of genotype × environment (G × E) interactions on their reactions to the disease, and identify genotypes with stability to the disease, using the Additive Main Effects and Multiplicative Interaction statistical model. Environments, genotypes and G × E interactions were highly significant (P < 0.01) for the disease. The G × E interactions accounted for 19.5% of the treatment sums of squares for CMD and influenced the relative ranking of genotypes across environments. The magnitude of the G × E interaction effect for CMD was larger than that of genotypes. Examination of the G × E interaction structure revealed specific areas where screening of cassava genotypes for resistance to CMD could be performed best. The study identified genotypes such as TMS 30001 and 63397 with resistance to CMD and CMD‐stable clone U/41044, which could be distributed to growers, and sites such as Ibadan and Ubiaja with high CMD severity for screening genotypes for reaction to CMD.     

Screening cassava for resistance to cassava mosaic disease by grafting and whitefly inoculation

Screening for cassava mosaic begomoviruses (CMBs)-resistance using grafting and whitefly inoculation was performed with local and improved cassava. The onset of symptom appearance and the evolution of Cassava mosaic disease (CMD) varied in function of genotypes and virus inoculation techniques used. Grafting position using cassava as scion or rootstock does not affect CMD display and evolution. No relation was established between the number of whiteflies feeding on each genotype and viral inoculation technique tested. Detopping of young leaves induces triggering effect on CMD expression. PCR and ELISA confirmed the EACMV-UG's preferential transmission by whitefly. Hypothesis of virus replication and cultivars's susceptibility were supported by virus increasing particles in infected cassava. Cultivars Mvuazi (TMSI 95/0528) and 96/1089A are suggested field immune to CMBs; Disanka (TMSI 95/0211), Yauma, Timolo, Bangi, Mahungu (TMS 92/297), Mvuama (TMS 83/138), Lueki (TMS 91/377) and Zizila (MV 99/0038) are CMD-resistant; whereas Ponjo, Lofiongi, Ngonga and Mboloko are susceptible. Our results showed that resistant genotypes may express CMD under high inoculum pressure such as grafting.     

Exploiting the Combination of Natural and Genetically Engineered Resistance to Cassava Mosaic and Cassava Brown Streak Viruses Impacting Cassava Production in Africa

Cassava brown streak disease (CBSD) and cassava mosaic disease (CMD) are currently two major viral diseases that severely reduce cassava production in large areas of Sub-Saharan Africa. Natural resistance has so far only been reported for CMD in cassava. CBSD is caused by two virus species, Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV). A sequence of the CBSV coat protein (CP) highly conserved between the two virus species was used to demonstrate that a CBSV-CP hairpin construct sufficed to generate immunity against both viral species in the cassava model cultivar (cv. 60444). Most of the transgenic lines showed high levels of resistance under increasing viral loads using a stringent top-grafting method of inoculation. No viral replication was observed in the resistant transgenic lines and they remained free of typical CBSD root symptoms 7 month post-infection. To generate transgenic cassava lines combining resistance to both CBSD and CMD the hairpin construct was transferred to a CMD-resistant farmer-preferred Nigerian landrace TME 7 (Oko-Iyawo). An adapted protocol allowed the efficient Agrobacterium-based transformation of TME 7 and the regeneration of transgenic lines with high levels of CBSV-CP hairpin-derived small RNAs. All transgenic TME 7 lines were immune to both CBSV and UCBSV infections. Further evaluation of the transgenic TME 7 lines revealed that CBSD resistance was maintained when plants were co-inoculated with East African cassava mosaic virus (EACMV), a geminivirus causing CMD. The innovative combination of natural and engineered virus resistance in farmer-preferred landraces will be particularly important to reducing the increasing impact of cassava viral diseases in Africa.     

Incidence of Three Major Cassava Diseases on Local Susceptible Cassava Cultivars at Three Planting Dates

Incidence of African cassava mosaic (ACMD), cassava bacterial blight (CBB), and cassava anthracnose (CA) on local susceptible cassava cultivars planted in December, February, and April was investigated. December cassava planting had a higher incidence of ACMD, CBB, and CA diseases compared with February and April plantings. CA seemed to be more prevalent at an older stage (8 months old) of plant growth. April planting had significantly lower incidence of major cassava diseases, and higher mean tuber, yield than the same cassava cultivars planted earlier. The length of exposure of cassava plants to the inocula and/or to the vector during the rainy season determines the occurrence of diseases. In the absence of resistant cassava cultivars and through, partial disease escape of susceptible cultivars from ACMD, CBB, and CA, planting the same cultivars in April can still produce a profitable cassava crop.     

Spatial and Temporal Spread of Cassava Mosaic Virus Disease in Cassava Grown Alone and when Intercropped with Maize and/or Cowpea

The spread of cassava mosaic disease (CMD) and populations of the whitefly vector (Bemisia tabaci) were recorded in cassava when grown alone and when intercropped with maize and/or cowpea. The trials were planted under conditions of high inoculum pressure in 1995 and 1996 at a site in the lowland rainforest zone of southern Cameroon. In the 1995 experiment, the maize and cowpea intercrops reduced the final incidence of CMD in the cassava cvs. Dschang White and Dschang Violet, but not in the more resistant cv. Improved. In the 1996 experiment with cv. Dschang Violet, the maize and cowpea intercrops grown alone or together decreased adult whitefly populations on cassava by 50% and CMD incidence by 20%. The monomolecular population growth model generally provided the best fit for disease progress. Areas under the disease progress curves (AUDPCs) and incidences expressed as multiple infection units were significantly (P<0.05) less for cassava intercropped with maize and/or cowpea than in cassava alone; times to 50% CMD incidence were significantly (P<0.05) longer in all intercrop systems. In 1995 the basic infection rates (r) were similarly low (0.010 per month) in the moderately resistant cv. Dschang Violet intercropped with maize and in all treatments in the more resistant cv. Improved. By contrast, rates were significantly higherfor cv. Dschang Violet alone or with cowpea and in all treatments for the less resistant cv. Dschang White (0.030–0.060). In 1996, r values in cassava grown alone (0.077) were significantly larger (P<0.05) than in the other cropping systems (0.042–0.052). There were no significant differences in the symptom severity in the different cropping systems. Disease foci were isodametric and more compact in plots containing cowpea than in other cropping systems.     

In vitro standardisation of resistance screening methods in cassava against tuber rot disease

Cassava tuber rot caused by Phytophthora palmivora in growing regions of Tamil Nadu and Kerala, is causing yield loss up to 80%. In the present study, resistance reactions of 10 cassava cultivars were analysed on leaf, stem and tuberous roots by artificial inoculation method in search of a suitable in vitro resistant screening method. Leaf and tuber analysis showed positive correlation (0.883) but the stem-based results showed negative correlation with leaf and tuber analysis. The analysis exhibited the susceptibility of the cassava cultivars against P. palmivora. Leaf analysis was superior in discriminating even small variations in resistance reactions than tuber analysis. The cultivar Sree Padmanabha showed higher resistance than other cultivars and the level of resistance in a cultivar is heritable which could be helpful in breeding programme. Based on the results it can be concluded that leaves of cassava could be used for screening resistance in the host and also in analysing the virulence of the isolate. This is the first report on screening the resistance in cassava cultivars against root rot caused by P. palmivora.     

The effects of cassava mosaic virus disease on yield and compensation in mixed stands of healthy and infected cassava

The effects of cassava mosaic virus disease (CMD) on yield in fully and partly infected stands of cassava were investigated in field trials in Uganda in 1990-91 and 1991-92. Three cultivars (Ebwanateraka, Bao and Bukalasa 1 l), each at three levels of cutting infection (O%, 50% and 100%) and harvested 510 and 15 months after planting (MAP) were used in a randomised block design with split-split plots and four replicates. Moreover, yield and growth data for individual infected and uninfected plants were considered in relation to the health status of their nearest neighbours. In each experiment, fresh tuberous root yields of plants from 100% infected plots gave sigdicantly lower yields than those from 0% or 50% infected plots at each harvest date and the losses were greatest in cv. Bao. Yields of plants from 0% and 50% plots for each of the three cultivars were not significantly different, 10 and 15 MAP. The loss in yield differed between cultivars and harvest dates. Fresh stem, leaf and root yields and the number of tuberous roots were influenced by the health status of the plants harvested and that of their nearest neighbours. Uninfected plants surrounded by infected ones had more roots and heavier total fresh root, stem and leaf weights than those surrounded by uninfected ones. Overall, 26% and 42% compensation was recorded in 1990-91 and 1991-92, respectively. The effects of CMD on cassava production and of compensation in mixed stands of infected and uninfected plants are discussed, especially in relation to control strategies such as roguing.     

Transcriptional Response of Virus-Infected Cassava and Identification of Putative Sources of Resistance for Cassava Brown Streak Disease

Cassava (Manihot esculenta) is a major food staple in sub-Saharan Africa, which is severely affected by cassava brown streak disease (CBSD). The aim of this study was to identify resistance for CBSD as well as to understand the mechanism of putative resistance for providing effective control for the disease. Three cassava varieties; Kaleso, Kiroba and Albert were inoculated with cassava brown streak viruses by grafting and also using the natural insect vector the whitefly, Bemisia tabaci. Kaleso expressed mild or no disease symptoms and supported low concentrations of viruses, which is a characteristic of resistant plants. In comparison, Kiroba expressed severe leaf but milder root symptoms, while Albert was susceptible with severe symptoms both on leaves and roots. Real-time PCR was used to estimate virus concentrations in cassava varieties. Virus quantities were higher in Kiroba and Albert compared to Kaleso. The Illumina RNA-sequencing was used to further understand the genetic basis of resistance. More than 700 genes were uniquely overexpressed in Kaleso in response to virus infection compared to Albert. Surprisingly, none of them were similar to known resistant gene orthologs. Some of the overexpressed genes, however, belonged to the hormone signalling pathways and secondary metabolites, both of which are linked to plant resistance. These genes should be further characterised before confirming their role in resistance to CBSD.     

Temporal spread of cassava mosaic virus disease in a range of cassava cultivars in different agro-ecological regions of Uganda

The spread of cassava mosaic disease (CMD) in a range of cassava cultivars was studied in experiments and on-farm trials in different agro-ecological regions of Uganda in 1989–1990 and 1990–1991. No spread occurred in either experiment at the southernmost site near Kampala, but there was considerable spread at the four sites elsewhere and also in the on-farm trials in Luwero district. There were significant differences in the final incidence of disease between locations and between cultivars at each location. Where spread occurred it was more rapid in the Ugandan cvs Ebwanateraka, Senyonjo and Bao than in four of the five improved TMS cultivars introduced from Nigeria. These usually showed an apparent decline in incidence of CMD after reaching maxima 4 to 8 months after planting (MAP). The areas under the disease progress curves (AUDPCs) differed significantly between locations and cultivars and were less for cvs TMS 30572, TMS 30395, TMS 30337 and TMS 60142 than for cvs Ebwanateraka, Senyonjo, Bao and TMS 30786. Overall, the mean AUDPCs were greatest at Migyera in Luwero district in 1989–1990 and at Kagando in Kasese district in 1990–1991. They were significantly less at Mubuku in Kasese district in 1989–1990 than at the other two experimental sites where spread occurred. Adult whitefly vector populations were highest at Migyera and Kagando in the 1989–1990 and 1990–1991 trials, respectively, and they were higher on cvs Bao, Ebwanateraka and TMS 30786 than on other varieties. Mean numbers of adults increased until 3–5 MAP and then declined, but CMD incidence increased progressively to reach maxima at or near crop maturity. Locations with the largest numbers of adults also had a relatively high incidence of CMD. Symptoms of CMD were usually more severe on cvs Ebwanateraka, Bao and Bukalasa 11 than on the TMS cultivars, on which symptoms remained slight throughout growth and usually decreased from 5 MAP. The differences between sites, the resistance of the cultivars and the relationship between CMD incidence and whitefly populations are discussed.     

Application of a Simplified Molecular Protocol to Reveal Mixed Infections with Begomoviruses in Cassava

Samples of cassava leaves exhibiting severe symptoms of cassava mosaic disease (CMD) were collected with the PhytoPASS kit in fields surrounding the city of Bujumbura (Burundi). These materials were then sent to Belgium for polymerase chain reaction determination of the CMD begomoviruses inducing the observed symptoms. Different pairs of specific primers were used to amplify DNA sequences specific to African cassava mosaic virus (ACMV), East African cassava mosaic virus (EACMV), East African cassava mosaic Cameroon virus (EACMCV), East African cassava mosaic Malawi virus (EACMMV), East African cassava mosaic Zanzibar virus (EACMZV), the Uganda variant of East African cassava mosaic virus (EACMV-UG) and South African cassava mosaic virus (SACMV). It was revealed that mixed infections were prevailing in the analyzed materials. Most of the samples submitted to this analysis were found to be co-infected by three different begomoviruses (ACMV + EACMV + EACMV-UG). The so revealed mixed infections could explain the high severity of CMD symptoms noticed on cassava in the region of Bujumbura while the diversity within the CMD causal agents illustrates the importance to take this parameter into consideration for a successful use of plant genetic resistance to control the disease.     

Cassava leaf harvesting as vegetables; a cause of vulnerability of cassava plant to cassava mosaic disease and eventual yield reduction: Ernte von cassavablättern als gemüse; eine ursache für die anfälligkeit der cassavapflanze für die cassava-mosaikkrankheit und für spätere ertragseinbußen

The consequence of harvesting young leaves of cassava as vegetable on the vulnerability of the crop to cassava mosaic disease (CMD) and on storage root yield was investigated using 30 cassava genotypes planted in IITA fields located in the humid forest (Port Harcourt?:?Onne), forest-savannah transition (Ibadan), southern guinea savannah (Mokwa) and northern guinea savannah (Zaria) agroecologies in Nigeria. Tender apical leaves and shoots of the cassava genotypes were removed from forty plants per cassava genotype with the same number of plants considered as control. Whitefly infestation, disease incidence (DI) and symptom severity (ISS) of the disease were assessed at monthly interval for six months and also at the ninth month after planting (MAP). Yield reduction due to this treatment was calculated as percentage harvest index (HI). Whitefly population fluctuated throughout the period of observation at all locations with higher population obtained generally for treated plants compared to control plants. Sprouting leaves of some treated genotypes were observed with severe mosaic symptoms, while corresponding control showed no mosaic symptoms. Contrarily, no remarkable difference was observed in Zaria between the mean ISS of treated and control cassava genotypes. There was a highly significant difference (P?<?0.01) in DI and ISS among cassava genotypes across all locations. Also, there was a highly significant interaction (P?<?0.01) in symptom severity between location (loc) and genotype, genotype and treatment (trt), loc and trt. Interaction between loc, genotypes and trt with regard to DI was highly significant at 2, 3 and 4 MAP, while with ISS, the interaction was highly significant all through the counting period. There was a positive relationship between DI and ISS on plants of genotypes 96/1039 and ISU. The percentage HI (27.4) of treated plants of genotype 95/0166 in Ibadan was remarkably lower than the value obtained for corresponding control (41.9) plants. Also, sharp distinction in% HI of treated (39.5) and control (43.8) ISU was observed in Onne with their respective ISS values as 3.7 and 3.2. Therefore, harvesting tender apical leaves and shoots of cassava as vegetables should be discouraged as it increases the severity of CMD infection in the regenerating shoots of cassava with attendant storage root yield reduction.     

The Aetiology of Vascular Discoloration in Cassava Roots after Harvesting: Development of Endogenous Resistance in Stored Roots

The susceptibility of cassava roots, Manihot esculenta Crantz, to vascular discoloration beneath two types of injury site, transverse cuts and periderm injuries, was compared for freshly harvested and stored roots. Susceptibility beneath transverse cuts changed rapidly, so that roots stored at ambient temperature for 5–9 days were largely resistant to vascular discoloration beneath this type of injury. Susceptibility to localized deterioration beneath periderm injuries changed more slowly, but significant decreases were observed in roots stored at ambient temperature for 10–16 days. Changes in susceptibility were observed in all cultivars tested, seven in Colombia and one in Jamaica. These changes were retarded but not prevented by storage at 2°C and by storage in sealed polyethylene bags. Pruning plants 1 to 3 weeks prior to harvesting, which has been shown to reduce the rate of post-harvest deterioration of roots, was also found to reduce the susceptibility of roots to vascular discoloration beneath injuries made immediately after harvesting. Water loss through injuries caused a respiratory response as well as vascular discoloration. This respiratory response was as large in stored (resistant) roots as in freshly harvested (susceptible) ones. The potential of cassava roots to develop endogenous resistance to vascular discoloration either before or after harvesting is discussed in relation to the problems of storage of harvested cassava roots.     

The evaluation of rhizomania resistant sugar beet for the UK

Sugar beet rhizomania disease, caused by Beet necrotic yellow vein virus and transmitted by the soil‐borne parasite Polymyxa betae, was first recorded in the UK in 1987. Recently, breeding lines and cultivars with partial resistance to the virus derived from the ‘Holly’ source of resistance have been developed and their suitability for use under UK conditions is explored in this paper. Virus multiplication in the roots of resistant lines exposed to severe disease pressure in glasshouse tests, when quantified by ELISA, was less than one third of that in susceptible controls. More recently developed resistant lines had a lower virus content, on average, largely due to a reduced frequency of susceptible individuals. There was no evidence for resistance to the vector, P. betae, in virus resistant lines. However, the proportion of viruliferous P. betae resting spores in the roots, estimated using the most probable number (MPN) technique, was reduced by at least one third in resistant lines compared with the most susceptible control. A novel line, containing an additional gene to that in ‘Holly’, was the most effective, reducing the infection level to 3% of that in the susceptible control. In two field experiments on severely infested sites, the rate of infection of a resistant line, when assessed by ELISA, was reduced by half compared with a susceptible cultivar and sugar yields of resistant lines were consistently 2–3 times higher than those of susceptible cultivars. In 41 trials on rhizomania‐free sites, several recently introduced resistant lines exhibited sugar yields and agronomic performance comparable to that of three selected high yielding, susceptible cultivars. Results are discussed in relation to the specific UK requirements for rhizomania resistant cultivars. One resistant line, Beta 805 (cv. Concept), fulfilled the requirements for widespread use to control the disease.     

Loss of CMD2‐mediated resistance to cassava mosaic disease in plants regenerated through somatic embryogenesis

Cassava mosaic disease (CMD) and cassava brown streak disease (CBSD) are the two most important viral diseases affecting cassava production in Africa. Three sources of resistance are employed to combat CMD: polygenic recessive resistance, termed CMD1, the dominant monogenic type, named CMD2, and the recently characterized CMD3. The farmer‐preferred cultivar TME 204 carries inherent resistance to CMD mediated by CMD2, but is highly susceptible to CBSD. Selected plants of TME 204 produced for RNA interference (RNAi)‐mediated resistance to CBSD were regenerated via somatic embryogenesis and tested in confined field trials in East Africa. Although micropropagated, wild‐type TME 204 plants exhibited the expected levels of resistance, all plants regenerated via somatic embryogenesis were found to be highly susceptible to CMD. Glasshouse studies using infectious clones of East African cassava mosaic virus conclusively demonstrated that the process of somatic embryogenesis used to regenerate cassava caused the resulting plants to become susceptible to CMD. This phenomenon could be replicated in the two additional CMD2‐type varieties TME 3 and TME 7, but the CMD1‐type cultivar TMS 30572 and the CMD3‐type cultivar TMS 98/0505 maintained resistance to CMD after passage through somatic embryogenesis. Data are presented to define the specific tissue culture step at which the loss of CMD resistance occurs and to show that the loss of CMD2‐mediated resistance is maintained across vegetative generations. These findings reveal new aspects of the widely used technique of somatic embryogenesis, and the stability of field‐level resistance in CMD2‐type cultivars presently grown by farmers in East Africa, where CMD pressure is high.     

AFLP analysis of African cassava (Manihot esculenta Crantz) germplasm resistant to the cassava mosaic disease (CMD)

Amplified fragment length polymophism was assessed in 20 land races and nine elite lines of cassava from Africa, resistant and susceptible to the cassava mosaic disease (CMD). Eleven accessions from a representative core collection from Latin America, previously studied by AFLPs, were included as a reference. AFLP data from all accessions was analyzed by both the unweighted pair group mean average (UPGMA) and multiple cluster analysis (MCA) methods of analysis. Genetic differentiation between clusters and the coefficient of genetic differentiation was also calculated. Results reveal a genetic divergence between African and Latin American accessions, although some overlap was found between them. African land races resistant to CMD, were also found to be genetically differentiated from susceptible land races and from resistant elite lines. AFLP analysis identified a considerable number of duplicates in the African accessions, suggesting a sizeable percentage of redundancy. A unique AFLP fragment, found in a relatively high frequency in African accessions, but absent in the Latin American accessions, was found to be associated with branching pattern by QTL mapping in an F1 progeny derived from African and Latin American parents. The likely source and the utility of the unique AFLP fragment in understanding the processes of genetic divergence in Africa is discussed. Received: 15 May 1999 / Accepted: 28 July 1999     

Rapid screenhouse assessment of bacterial blight in cassava genotypes in Nigeria

Bacterial blight disease caused by Xanthomonas axonopodis pv. manihotis (Berthet-Bondar) Dye was assessed in 11 artificially inoculated cassava genotypes in a screenhouse. Disease progress was estimated at intervals of 3 days by measuring the length of necrotic lesions on stems and leaves, as well as estimating the average disease score and area under disease progress curve (AUDPC). Based on the average disease scores, cassava genotypes 30572, TME 1, TME 7 and TME 9 were classified as resistant to bacterial blight, genotypes 4(2)1425, TME 2, TME 4 and TME 12 were tolerant while cassava genotypes 30001, TME 3, and TME 28 were susceptible. Direct correlations, statistically significant at p < 0.05, were obtained between stem necrosis, leaf necrosis, average disease scores and AUDPC in the 11 cassava genotypes. Screenhouse experiments afford rapid assessment of resistance status of cassava genotypes to bacterial blight in Nigeria.