Sweet Potato Virus Disease Research Articles

Synergistic Interaction of Sweet potato chlorotic stunt virus (Crinivirus) with Carla-, Cucumo-, Ipomo-, and Potyviruses Infecting Sweet Potato.

Co-infection of Sweet potato chlorotic stunt virus (SPCSV, genus Crinivirus) with Sweet potato feathery mottle virus (SPFMV, genus Potyvirus) results in sweet potato virus disease (SPVD), a synergistic disease that is widely distributed in the sweet potato (Ipomoea batatas) growing regions of the world. Since both SPCSV and SPFMV are common and often detected as part of multiple co-infections of severely diseased plants, the occurrence of synergistic interactions with other viruses was investigated. Data from this study show that SPCSV, but not SPFMV, can cause synergistic diseases in sweet potato with all viruses tested, including members of the genus Potyvirus (Sweet potato latent virus, Sweet potato mild speckling virus), Ipomovirus (Sweet potato mild mottle virus), Cucumovirus (Cucumber mosaic virus), and putative members of the genus Carlavirus (Sweet potato chlorotic fleck virus and C-6 virus). The synergism was expressed as an increase in the severity of symptoms, virus accumulation, viral movement in plants, and as an effect on yield of storage roots. The presence of a third different virus in plants affected with SPVD increased the severity of symptoms even further compared with SPVD alone. There was a positive correlation between increase in virus accumulation and symptom expression in double and triple SPCSV-associated co-infections. The epidemiological implications of the results are discussed.

Viruses and virus-like diseases affecting sweet potato subsistence farming in southern Tanzania

A survey for sweet potato viruses was conducted in 32 farmer’s fields in Mbinga (20 fields) and Songea (12 fields) districts of the Ruvuma region in Southern Tanzania. Plants showing virus-like symptoms were observed in 25 (78%) out of 32 fields examined. Sweet potato virus disease (SPVD) incidence was low in Mbinga district (16.7%) on average and ranged from 3 to 50% while in Songea district incidence averaged 33% and ranged from 3 to 100%. SPVD incidence difference between the two districts was not statistically significant (P > 0.05) except among sweet potato cultivars (P< 0.05). Sweet potato virus disease severity significantly (P < 0.001) varied between the district with Mbinga having the lowest (2.38 ± 0.2) and Songea the highest (3.26 ± 0.12) mean severity score. SPVD severity score also significantly varied among sweet potato cultivars (P < 0.001) with cultivar ‘‘Jeshi’’ displaying the most severe symptoms (4.2 ± 0.37 severity score) and ‘Wino’ and ‘Madaresalama expressing the mildest symptoms (2.0 ± 0.00). Twenty foliar samples from infected plants were tested serologically for Sweet potato chlorotic stunt virus (SPCSV), Sweetpotato feathery mottle virus (SPFMV), Sweet potato mild mottle virus (SPMMV), Sweet potato chlorotic fleck virus (SPCFV), Sweet potato calico-like virus (SPCaLV), Sweet potato mild speckling virus (SPMSV), C-6 (a new flexious rod virus), Sweet potato latent virus (SwPLV), Sweet potato virus G (SPVG) andCucumber mosaic virus (CMV) using Nitrocelullose Membrane ELISA (NCM-ELISA). SPCFV was the most common virus detected followed by SPCSV. SPVG was detected in seven samples representing the first report of its occurrence in Tanzania. SwPLV, C-6, CMV and SPCaLV were not detected. Whitefly population was low and aphids were rarely found in most of the fields. Orange-fleshed sweet potato cultivars grown in the surveyed areas also displayed SPVD symptoms with high incidence and severity. These results indicate that SPVD situation in Tanzania is not uniform in the major sweet potato growing areas and the significance of this virus diversity needs to be investigated. Key words: Sweet potato, sweet potato virus disease, NCM-ELISA, incidence, Tanzania

First Report of Sweet potato chlorotic stunt virus, a Component of Sweetpotato Virus Disease, in North Carolina.

Sweet potato chlorotic stunt virus (SPCSV) is the whitefly-transmitted component of the sweet potato virus disease (SPVD), a devastating disease originally described in Africa (4). Two isolates designated as G-01 and T-03 were obtained in North Carolina in July 2001 and October 2003, respectively, from plants of cv. Beauregard exhibiting symptoms typical of SPVD, including stunting, leaf narrowing and distortion, vein clearing, and chlorotic mosaic. Sap extract from symptomatic plants tested positive for SPCSV by nitrocellulose immuno-dot blot, using monoclonal antibodies specific for SPCSV obtained from the International Potato Center. Total RNA was extracted from 100 mg of symptomatic leaf tissue by using the PureLink Total RNA Purification System Kit from Invitrogen (Carlsbad, CA) with a minor modification (adding 2% PVP-40 and 1% 2-mercaptoethanol to the extraction buffer) (1). Results were confirmed by reverse transcription (RT)-PCR using primers CP1 and CP3 and HSP70-A/HSP70-B (2), corresponding to the capsid protein and 'heat shock' protein genes, respectively. HSP70 amplicons were cloned using the TOPO TA Cloning Kit (Invitrogen) and sequenced. At the nucleotide level, viral sequences from clones from both isolates were an average 99.4% similar to West Africa and 77.9% to East Africa sequences of SPCSV from Genbank (1). Although the isolates were collected from different fields, viral sequences generated from clones for T-03 and G-01 differed by only six nucleotides and were identical at the amino acid level. The neighbor-joining phylogenetic tree constructed using the HSP70 gene fragment (39 nt) delineated two major clusters with two subpopulations each: Cluster 1, "East Africa", consisted of East Africa and Peru subpopulations; Cluster 2, "West Africa", consisted of Argentina-Brazil and USA-West Africa subpopulations (1). In addition, SPCSV isolates from East Africa and West Africa clusters were sufficiently distant phylogenetically to suggest that they may correspond to two different criniviruses, with an average similarity between the populations of 78.14% and an average within the populations above 89%. Hudson's tests confirmed the presence of genetically distinct SPCSV groups with high statistical significance (1). Two groups (Peru and East Africa) were differentiated in the East Africa cluster, and three groups (Argentina-Brazil, USA, and West Africa) were differentiated in the West Africa cluster, suggesting that the USA population is not a recent introduction. Although SPCSV was previously reported in the United States, the source was a single accession of cv. White Bunch from the USDA Sweetpotato Germplasm Repository (3). Sweet potato feathery mottle virus (SPFMV) (family Potyviridae, genus Potyvirus), the other component of SPVD, was also detected in both cultivars. To our knowledge, this is the first report of SPCSV in sweetpotato fields in the United States.

Localization of Sweet potato chlorotic stunt virus (SPCSV) in synergic infection with Potyviruses in sweet potato

Among diseases reported worldwidely for sweet potato (Ipomoea batatas (L) Lam) crop, one of the most frequent is the Sweet potato virus disease (SPVD), caused by sweet potato chlorotic stunt virus (SPCSV) and sweet potato feathery mottle virus (SPFMV) co-infection. In Argentina, there exists the sweet potato chlorotic dwarf (SPCD), a sweet potato disease caused by triple co-infection with SPCSV, SPFMV and sweet potato mild speckling virus (SPMSV). Both diseases cause a synergism between the potyviruses (SPFMV and SPMSV) and the crinivirus (SPCSV). Up to date, studies carried out on the interaction among these three viruses have not described their localization in the infected tissues. In single infections, virions of the crinivirus genus are limited to the phloem while potyviral virions are found in most tissues of the infected plant. The purpose of this work was to localize the heat shock protein 70 homolog (HSP70h), a movement protein for genus crinivirus, of an Argentinean SPCSV isolate in its single infection and in its double and triple co-infection with SPFMV and SPMSV. The localization was made by in situ hybridization (ISH) for electron microscopy (EM) on ultrathin sections of sweet potato cv. Morada INTA infected tissues. The results demonstrated that viral RNA coding HSP70h is restricted to phloem cells during crinivirus single infection, while it was detected outside the phloem in infections combined with the potyviruses involved in chlorotic dwarf disease.

Sweet potato feathery mottle virus is the casual agent of sweetpotato virus disease in Italy

Mungbean yellow mosaic India virus (MYMIV), a bipartite begomovirus, is prevalent on several legume crops in the Indian sub-continent (Pant et al. , 2001) and has previously been reported affecting mungbeans in Pakistan (Hussain et al ., 2004). Mothbean ( Vigna aconitifolia ) is grown in only in a few areas of the country, mostly as animal fodder. Recently, work on improvement has been initiated due to its potential both for animal and human nutrition. As a first step, seeds of available mothbean varieties were screened under field conditions. All varieties exhibited a severe yellow mosaic, along with leaf curling on some varieties. Based on symptoms the involvement of MYMIV was suspected. To confirm the identity of the virus, total DNA was extracted from samples of infected plants, resolved in agarose gels and blotted to nylon membranes. A full length MYMIV DNA A clone isolated from mungbean (unpublished results) was used as a probe in Southern hybridisation and detected bands typical of geminiviruses in all samples with symptoms, but not those without; confirming the association of a begomovirus with the disease. To further confirm the identity of the virus, primers specific for the DNA B-encoded nuclear shuttle protein (NSP) gene of MYMIV (Hussain et al ., 2004) were used in PCR. These resulted in amplification of an approximately 800 bp fragment, that was cloned and sequenced. The complete 771 bp sequence of the NSP gene (accession number AM233490) showed 95–92% nucleotide sequence identity (94– 92% amino acid similarity) to the NSP gene of other MYMIV isolates. These results confirm the association of MYMIV with yellow mosaic disease of mothbean and is the first report of MYMIV infecting this species.

Interactions Among Sweet potato chlorotic stunt virus and Different Potyviruses and Potyvirus Strains Infecting Sweetpotato in the United States.

Sweet potato virus disease (SPVD) is the most serious viral disease of sweetpotato globally. This disease is caused by the synergistic interaction between the aphid-transmitted potyvirus Sweet potato feathery mottle virus (SPFMV) and the whitefly-transmitted crinivirus Sweet potato chlorotic stunt virus (SPCSV). In sweetpotato, titers of SPFMV have been shown to be significantly enhanced when coinfecting with SPCSV. In this study, the effect of SPCSV on titers of different potyviruses and potyvirus strains infecting sweetpotato in the United States was investigated using real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). No significant difference was observed in titers of the russet crack strain of SPFMV (SPFMV-RC), Ipomoea vein mosaic virus (IVMV), and Sweet potato virus G (SPVG) between single and mixed infections. Titers of all potyviruses and potyvirus strains evaluated were enhanced in the presence of SPCSV, suggesting that a conserved mechanism may underlie these interactions. Titers of the common strain of SPFMV (SPFMV-C), which did not cause SPVD-like symptoms when coinfecting with SPCSV, were also significantly enhanced in the presence of SPCSV. Furthermore, titers of SPCSV were found to be lower in treatments involving pairwise infections compared with plants infected by SPCSV alone. The degree of potyvirus titer enhancement did not correspond to the severity of symptoms observed in certain treatments involving pairwise infections.

The Effect of Sweet Potato Virus Disease and its Viral Components on Gene Expression Levels in Sweetpotato

Sweet potato virus disease (SPVD) is the most devastating disease of sweetpotato [Ipomoea batatas (L.) Lam.] globally. It is caused by the co-infection of plants with a potyvirus, sweet potato feathery mottle virus (SPFMV), and a crinivirus, sweet potato chlorotic stunt virus (SPCSV). In this study we report the use of cDNA microarrays, containing 2765 features from sweetpotato leaf and storage root libraries, in an effort to assess the effect of this disease and its individual viral components on the gene expression profile of I. batatas cv. Beauregard. Expression analysis revealed that the number of differentially expressed genes (P &lt; 0.05) in plants infected with SPFMV alone and SPCSV alone compared to virus-tested (VT) plants was only 3 and 14, respectively. However, these findings are in contrast with SPVD-affected plants where more than 200 genes were found to be differentially expressed. SPVD-responsive genes are involved in a variety of cellular processes including several that were identified as pathogenesis- or stress-induced.

Virus Disease Incidences of Sweet Potatoes in Korea

In 2003, a survey of sweet potato virus disease was carried out in seed boxes as well as in various sweet potato fields. Virus infection rate was 5~100% and 100% at seed boxes and fields, respectively. No relationship of the disease incidence and severity was observed between sweet potato cultivating areas and cultivars. A total of 179 samples were collected and analyzed based on serological, electron microscopic and molecular properties. Field-grown sweet potatoes were examined to inspect 8 different viruses using NCM-ELISA, resulting that 30% of sweet potato was infected by one virus, whereas 70% was by more than 2 viruses. However, RTPCR using primers selected for seven viruses, such as Sweet potato feathery mottle virus (SPFMV) revealed that of one-hundred seventy-nine tested; 71 of SPFMV, 29 of SPGV, 19 of SPFMV+SPGV, 1 of SPFMV+ SwPLV, 1 of SPFMV+SPLCV, 2 of SPFMV+SPGV+ SwPLV, 6 of SPFMV+SPGV+SPLCV, 2 of SPFMV+ SPGV+SwPLV+SPLCV and 48 of unknown viruses were identified from the field samples. In root, viral diseases were severer in Yeoju than in Mokpo Experiment Station and infection rate was much different depending on sweet potato cultivars.

Detection and Distribution of Viruses Infecting Field-grown Sweetpotato in East Africa

Sweetpotato is an important staple food crop in Sub-Saharan Africa, with production being concentrated in East Africa, particularly around Lake Victoria. Productivity of the crop is greatly constrained by viral diseases. Four main viruses have consistently been detected from various surveys done in the region viz., sweetpotato feathery mottle virus (SPFMV), sweetpotato chlorotic stunt virus (SPCSV), sweetpotato mild mottle virus (Sp.m.MV), and sweetpotato chlorotic fleck virus (SPCFV). The most severe symptoms have been caused by co-infection with SPCSV and SPFMV, resulting in the synergistic sweetpotato virus disease (SPVD). Some local sweetpotato genotypes have been reported to recover from, or have localized distribution of SPVD, suggesting that the disease is not fully systemic. This has led to the suggestion that uninfected cuttings may be obtained from previously infected plants. Experiments were set to determine the possibility of obtaining cuttings long enough for propagation that are free from virus infection. This would form a basis for recommending to the local small-holder farmers of a way to reduce losses due to the disease. Field-grown sweetpotato vines were cut into three pieces (15, 15–30, and &gt;30 cm from the apex) and tested for SPCSV and SPFMV. Nine genotypes were selected from a group of 21 local clones and used for this study. The two viruses were equally present in all the three sections of infected vines, indicating that it is not easy to obtain a virus-free cutting for field propagation from an infected vine. Virus assays in the past has mainly been limited to the use of serological methods. Use of PCR resulted in detection of begomoviruses infecting sweetpotatoes for the first time in the region.

AN UPDATE ON EVALUATION OF VIRUS STATUS IN THE HEIRLOOM SWEETPOTATO GERMPLASM MATERIALS WITH REAL-TIME PCR TECHNOLOGY

Sweet potato virus disease (SPVD) is the most devastating virus disease on sweetpotato [Ipomoea batatas (L.) Lam] world wide, especially in East Africa. However, weather it is present in the U.S. is unknown. SPVD is caused by co-infection of sweetpotato feathery mottle virus (SPFMV) and sweetpotato chlorotic stunt virus (SPCSV). Presence of two other potyviruses, sweetpotato virus G (SPVG) and Ipomoea vein mosaic virus (IVMV) has also been confirmed in the U.S. Sweet potato leaf curl virus (SPLCV), a whitefly (Bemisia tabaci) transmitted Begomovirus, also has the potential to spread to commercial sweetpotato fields and poses a great threat to the sweetpotato industry. The U.S. collection of sweetpotato germplasm contains about 700 genotypes or breeding lines introduced from over 20 different countries. Newly introduced sweetpotato germplasm from foreign sources are routinely screened for major viruses with serology and graft-transmission onto indicator plants (Ipomoea setosa). However, a large portion of this collection including heirloom cultivars or old breeding materials has not been systemically screened for these major sweetpotato viruses. In this study, a total of 69 so-called heirloom sweetpotato PI accessions were evaluated for their virus status. We used Real-time PCR to detect five sweetpotato viruses, including four RNA viruses (SPCSV, SPFMV, SPVG, and IVMV) and one DNA virus (SPLCV). A multiplex Real-time RT-PCR system was developed to detect three RNA viruses (SPFMV, SPVG, and IVMV). Preliminary data indicated that about 15% of these heirloom sweetpotato germplasm carried at least one of these viruses tested. Details on virus infection status will be presented.

VIRUS DISTRIBUTION IN FIELD GROWN SWEETPOTATO VINES IN AFRICA

Sweet potato virus disease (SPVD) is a major constraint to sweetpotato production in East Africa. The disease is a result of co-infection with sweet potato feathery mottle virus (SPFMV) and sweet potato chlorotic stunt virus (SPCSV). Some local sweetpotato genotypes have been reported to recover from, or have localized distribution of SPVD, suggesting that the disease is not fully systemic. This has led to the suggestion that uninfected cuttings may be obtained from previously infected plants. Experiments were set to determine the possibility of obtaining cuttings long enough for propagation that are free from virus infection. This would form a basis for recommending to the local small-holder farmers of a way to reduce losses due to the disease. Field grown sweetpotato vines were cut into three pieces (15, 15 to 30, and &gt;30 cm from the apex) and tested for SPCSV and SPFMV. Nine genotypes were selected from a group of 21 local clones and used for this study. The two viruses were equally present in all the three sections of infected vines, indicating that it is not easy to obtain a virus free cutting for field propagation from an infected vine.

First Report of a Begomovirus Infecting Sweetpotato in Kenya

Previous surveys for viruses in sweetpotatoes (Ipomoea batatas) in Africa did not assay for the presence of begomoviruses such as Sweet potato leaf curl virus (SPLCV), which have been found recently in the Americas and Asia. Symptomatic sweetpotato plants, including some with leaf curling symptoms similar to those observed in SPLCV-infected sweet-potato plants (2), were collected from a germplasm collection plot at Kakamega Research Station in Western Kenya during February 2005. Whiteflies, the vectors for begomoviruses, were observed in the same plots. Ipomoea setosa plants graft-inoculated with scions from the symptomatic sweetpotato developed leaf curl, leaf roll, interveinal chlorosis, and stunting symptoms similar to those caused by infection with SPLCV alone or in combination with Sweet potato feathery mottle virus. Total DNA was isolated from 10 I. setosa plants using the GenElute Plant Genomic DNA Kit (Sigma-Aldrich Inc., St. Louis, MO). Sweetpotato cuttings from 39 clones, selected from the Kenyan germplasm collection for their resistance or susceptibility to sweetpotato virus disease (SPVD), were sent to the Plant Germplasm Quarantine Office of USDA-ARS. The cuttings were planted in a greenhouse. Total DNA was extracted from sweetpotato leaves 1 month later using a cetyltrimethylammoniumbromide (CTAB) extraction method (1). Degenerate primers SPG1/SPG2, developed for PCR detection of begomoviruses (1), amplified a 912-bp DNA fragment from 3 of 10 DNA extracts from I. setosa and 5 of 39 sweetpotato plants held in quarantine. The primers anneal to regions of open reading frame (ORF) AC2 and ORF AC1 that are highly conserved in begomoviruses infecting sweetpotato. SPLCV-specific primers PW285-1/PW285-2 (2) amplified a 512-bp DNA fragment of ORF AC1 from seven samples (two from I. setosa and five from I. batatas). Amplicons from three independent PCR assays of two samples and single PCR assays of four additional samples were cloned into the pGEM-T Easy vector. Clone inserts were sequenced, and compared with sequences deposited in GenBank using the basic local alignment search tool (BLAST). Sequences were closely related to SPLCV (GenBank Accession No. AF104036) with nucleotide sequence identities varying from 93% (GenBank Accession No. DQ361004) to 97% (GenBank Accession No. DQ361005). The presence of the virus poses a challenge to the dissemination of planting materials in the region because begomovirus-infected plants often do not show symptoms. To our knowledge, this is the first report of a begomovirus infecting sweetpotato in Kenya or the East African Region.

SWEETPOTATO VIRUS DISEASES RESEARCH IN EAST AFRICA

Sweetpotato is an important staple food crop in Sub-Saharan Africa, with production being concentrated in East Africa, particularly around Lake Victoria. Productivity of the crop is greatly constrained by viral diseases. Four main viruses have consistently been detected from various surveys done in the region viz. sweet potato feathery mottle virus (SPFMV), sweet potato chlorotic stunt virus (SPCSV), sweet potato mild mottle virus (SPMMV), and sweet potato chlorotic fleck virus (SPCFV). Sweet potato caulimo-like virus (SPCaLV), sweet potato latent virus (SPLV), and cucumber mosaic virus (CMV) have also been detected though only in isolated cases. The most severe symptoms have been caused by co-infection with SPCSV and SPFMV, resulting in the synergistic Sweet potato virus disease (SPVD). Yield reductions due to virus infections have been estimated to be &gt;90% in very severe cases. Virus detection has mainly been limited to the use of serological methods. Some plants have been observed with symptoms resembling those caused by viruses, but do not react with available antisera, indicating that the plants could be infected with viruses that have not been described, or not tested in the region. Use of other detection techniques such as PCR may result in identification of more viruses in the region. This report gives a summary of our research efforts towards detection of other viruses present in the region, and identification of resistant germplasm.

Interactions between a crinivirus, an ipomovirus and a potyvirus in coinfected sweetpotato plants

Novel and severe symptoms of chlorosis, rugosity, leaf strapping and dark green islands, designated as sweetpotato severe mosaic disease (SPSMD), were caused by dual infection of Sweet potato mild mottle virus (SPMMV; Ipomovirus) and Sweet potato chlorotic stunt virus (SPCSV; Crinivirus) in three East African sweetpotato cultivars (Tanzania, Dimbuka and New Kawogo). The storage root yield was reduced by ∼80%, as compared with healthy plants under screenhouse conditions in Uganda. Plants infected with SPMMV or SPCSV alone showed nonsignificant or 50% yield reduction, respectively. SPCSV reduced resistance to SPMMV in sweetpotato, similar to the situation with resistance to Sweet potato feathery mottle virus (SPFMV; Potyvirus) that breaks down following infection with SPCSV, followed by development of sweet potato virus disease (SPVD). In single virus infections with SPMMV and SPFMV or their coinfection, cvs Tanzania and Dimbuka were initially systemically infected, displayed symptoms and contained readily detectable virus titres, but new leaves were symptomless with very low virus titres, indicating recovery from disease. In contrast, cv. New Kawogo remained symptomless and contained low SPMMV and SPFMV titres following graft inoculation. These moderate and high levels of resistance to SPMMV and SPFMV, respectively, were lost and cultivars succumbed to a severe disease following coinfection with SPCSV. The synergistic interactions increased titres of SPMMV and SPFMV RNA by ∼1000‐fold as quantified by real‐time PCR, whereas SPCSV titres were reduced twofold, indicating an antagonistic interaction. Coinfection with SPMMV and SPFMV caused no detectable changes in virus titres or symptom severity.

CHARACTERISTICS AND DIVERSITY IN SWEETPOTATO-INFECTING VIRUSES IN AFRICA

In Africa, seven viruses occurring singly or in combination have been identified in sweetpotato. The viruses include Sweet potato feathery mottle potyvirus (SPFMV), Sweet potato chlorotic stunt crinivirus (SPCSV), Sweet potato mild mottle ipoitiovirus (SPMMV), Sweet potato chlorotic fleck virus (SPCFV), Sweet potato virus II (SPVII), Sweet potato caulimo-like virus (SPCaLV) and Sweet potato virus G (SPVG). Out of these, the presence of SPFMV was detected in most countries surveyed, while SPCSV occurred in Nigeria, Gabon, Kenya, Uganda, Tanzania, Zambia, Madagascar and Egypt. SPCFV and SPMMV had limited distribution, mainly in East Africa. SPCaLV was only found in Uganda, SPVII in South Africa and SPVG in Ethiopia and Egypt. Molecular characterisation showed that most of the virus isolates have wide genetic variations, although some such as SPCSV are highly conserved. SPCSV is unique among these viruses because it has the ability to synergise others, therefore, is considered to be the most important virus responsible for the development of severe symptoms characteristic of sweetpotato virus disease, hence, it should be a major target for control.

Unravelling the genetic diversity of the three main viruses involved in Sweet Potato Virus Disease (SPVD), and its practical implications

SUMMARY Sweetpotato (Ipomoea batatas) is a widely grown food crop, in which the most important diseases are caused by viruses. Genetic variability of three widely distributed sweetpotato viruses was analysed using data from 46 isolates of Sweet potato feathery mottle virus (SPFMV), 16 isolates of Sweet potato mild mottle virus (SPMMV) and 25 isolates of Sweet potato chlorotic stunt virus (SPCSV), of which 19, seven and six isolates, respectively, are newly characterized. Division of SPFMV into four genetic groups (strains) according to phylogenetic analysis of coat protein (CP) encoding sequences revealed that strain EA contained the East African isolates of SPFMV but none from elsewhere. In contrast, strain RC contained ten isolates from Australia, Africa, Asia and North America. Strain O contained six heterogeneous isolates from Africa, Asia and South America. The seven strain C isolates from Australia, Africa, Asia, and North and South America formed a group that was genetically distant from the other SPFMV strains. SPMMV isolates showed a high level of variability with no discrete strain groupings. SPCSV isolates from East Africa were phylogenetically distant to SPCSV isolates from elsewhere. Only from East Africa were adequate data available for different isolates of the three viruses to estimate the genetic variability of their local populations. The implications of the current sequence information and the need for more such information from most sweetpotato-growing regions of the world are discussed in relation to virus diagnostics and breeding for virus resistance.

Studies on the physiological effects of viruses on sweet potato yield in Kenya

SummaryExperiments to determine the effect of the three viruses most common on Kenyan sweet potato varieties were carried out at the University of Nairobi Kabete farm. The sweet potato varieties were Bungoma, Kemb 10 and Ex‐Shimba hills which were either virus‐free or infected with Sweet potato feathery mottle virus (SPFMV), Sweet potato mild mottle virus (SPMMV), Sweet potato chlorotic stunt virus (SPCSV) or a combination of SPFMV and SPCSV. Canopy photosynthetically active radiation (PAR) interception and the relative chlorophyll content of the plants were determined. At harvest, marketable tuber yield, harvest index (HI), leaf area index (LAI) and specific leaf weight (SLW) were measured.Infection with single viruses caused no or mild symptoms and had no significant effect on chlorophyll content of leaves of any variety. Dual infection with SPFMV and SPCSV resulted in severe symptoms characteristic of sweet potato virus disease complex (SPVD) and significantly reduced chlorophyll content. Only SPCSV and SPCSV + SPFMV reduced PAR interception, the latter combination especially. Generally, infection with single viruses increased the SLW but a decrease occurred in plants dually infected with SPFMV and SPCSV. Whereas SPVD significantly reduced the harvest index (78%) and tuber yield (98%) in the three varieties, infection with single viruses caused a reduction only in the varieties Bungoma and Kemb 10. SPVD reduced yield through reducing leaf chlorophyll content, PAR interception, LAI and HI. The single virus infections possibly reduced assimilate translocation from leaves to tubers as is inferred from high SLW. Effects of viruses on physiological processes and yield are discussed.

First Report of Sweet potato chlorotic stunt virus and Sweet potato feathery mottle virus Infecting Sweet Potato in Spain.

Sweet potato chlorotic stunt virus (SPCSV), family Closteroviridae and Sweet potato feathery mottle virus (SPFMV), family Potyviridae are whitefly and aphid transmitted, respectively, which in double infections cause sweet potato virus disease (SPVD) that is a serious sweet potato (Ipomoea batatas Lam.) disease in Africa (2). During the past decade, sweet potato plants showing symptoms similar to SPVD have been observed in most areas of Spain. Nevertheless, not much information is available about the identity of the viruses infecting this crop in Spain. During the summer of 2002, sweet potato plants with foliar mosaic, stunting, leaf malformation, chlorosis, and ringspot symptoms were observed in several farms in Málaga (southern Spain) and Tenerife and Lanzarote (Canary Islands, Spain). Vine cuttings were collected from 21 symptomatic plants in Málaga and from eight plants on Lanzarote and six on Tenerife. Scions were grafted to the indicator hosts, Brazilian morning glory (I. setosa) and I. nil cv. Scarlett O'Hara. Three weeks after graft inoculations, all plants showed various degrees of mosaic, chlorosis, leaf malformation, and stunting. Four field collections (two from Málaga, one from Tenerife, and one from Lanzarote) with severe symptoms on I. setosa were selected for whitefly (Bemisia tabaci biotype Q) transmission experiments. Acquisition and transmission periods were 48 h. I. setosa was the acquisition host, and I. nil was the transmission host. For each isolate, groups of 10 whiteflies per I. nil plant were used. All I. nil plants used as transmission hosts with the four, field collections showed chlorosis and leaf malformation. Reverse-transcription polymerase chain reaction (RT-PCR) was performed on I. setosa (grafted with the four selected field collections) and I. nil plants (from the whitefly transmission experiments) with primers for the HSP70h gene of SPCSV. A 450-bp DNA fragment was obtained with all I. setosa and I. nil samples. Sequencing of the 450-bp DNA from two samples from Málaga yielded a nucleotide sequence with 98 to 99% similarity to the HSP70h gene of West African SPCSV isolates. Foliar samples from I. setosa, originally grafted with the 21 vine cuttings, were used for nitrocellulose membrane enzyme-linked immunosorbent assay (NCM-ELISA) testing with antiserum specific to SPFMV-RC (provided by J. Moyer, North Carolina State University, Raleigh). Positive control was sap extract from I. setosa that was infected with the common strain of SPFMV. Procedures for NCM-ELISA were as described (4). NCM-ELISA testing suggested that SPFMV was present in all samples. RT-PCR was conducted with degenerate primers POT1/POT2 (1). The nucleotide sequence that was amplified by these two primers spans part of the NIb protein and part of the coat protein gene of potyviruses. All samples yielded the expected 1.3-kb DNA. Sequencing of the RT-PCR products of two isolates from Malaga and sequence comparisons yielded nucleotide sequences with 97% similarity to two East African isolates (Nam 1 and Nam 3) of SPFMV (3). These results confirm the presence of SPCSV and SPFMV in sweet potato in Spain.

Control strategies for sweet potato virus disease in Africa

Sweet potato virus disease (SPVD), caused by dual infection with the whitefly-borne Sweet potato chlorotic stunt virus (SPCSV) and the aphid-borne Sweet potato feathery mottle virus (SPFMV), is the most serious disease of sweet potato in Africa. SPVD has been known there since at least the 1940s, although it took several decades to elucidate its aetiology. It occurs throughout Africa and is particularly prevalent in the Great Lakes region. Production of sweet potato is largely by resource-poor farmers, growing mostly local landraces and for home consumption and so control strategies need to be appropriate to these circumstances. Most high yielding and/or early maturing landraces in Uganda are susceptible to SPVD and most resistant landraces are low-yielding, forcing farmers to compromise between the conflicting requirements of large and/or early yields, and food security. Accordingly, two strategies were tested to avoid the disadvantages associated with such compromises. These were deploying high-yielding SPVD-resistant cultivars and phytosanitation practices to enable susceptible landraces to be grown successfully. In on-farm trials in Masaka and Rakai Districts of Uganda, some SPVD-resistant cultivars bred at Namulonge Agricultural and Animal Research Station (NAARI), in Wakiso District, out-yielded local landraces. Other trials at NAARI and at nearby farms showed that roguing diseased cuttings within 1 month of planting and isolation from diseased crops, even by as little as 15 m, can considerably decrease spread of SPVD to susceptible cultivars. This indicates that phytosanitation can protect desirable susceptible cultivars, even if adopted only locally. A dual approach of deploying both resistant varieties and phytosanitation provides farmers with a valuable increase in their choice of control strategies for SPVD.

Within-crop spread of sweet potato virus disease and the population dynamics of its whitefly and aphid vectors

Sweet potato virus disease (SPVD) is the most damaging disease of sweet potato ( Ipomoea batatas (L) Lam) in Uganda and results from dual infections of Sweet potato feathery mottle virus (SPFMV), vectored by aphids, and Sweet potato chlorotic stunt virus (SPCSV), vectored by whiteflies. For three cropping cycles, whiteflies ( Bemisia tabaci), aphids ( Myzus persicae and Aphis gossypii) and the spread of SPVD were monitored within and outside the sweet potato crop containing a central plot of SPVD-infected plants. There were similar catches of whiteflies on sticky traps placed on poles within the crop at different distances and directions around the infected plot. Outside the crop, there were also similar catches of whiteflies near (15 m) and far away (100 m) from the sweet potato crop. However, more whiteflies were trapped within than outside the crop. Most whiteflies were trapped close to the canopy. Aphids were rarely found on sweet potato plants, although they were commonly found on the traps. The number of aphids trapped inside the crop did not differ significantly from those trapped outside the crop. Consistently, a higher proportion of plants developed SPVD symptoms in the inner square than plants located further away from the infected plot. Number of infected plants correlated with trap catches of whiteflies only for the first season.